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DISCOVERING RETROVIRUSES
DISCOVERING RETROVIRUSES

Beacons in the Biosphere

Anna Marie Skalka

Cambridge, Massachusetts
London, England
2018
Copyright © 2018 by the President and Fellows of Harvard College
All rights reserved
Printed in the United States of Amer­i­ca
First Printing

Library of Congress Cataloging-­in-­Publication Data

Names: Skalka, Anna Marie, author.
Title: Discovering retroviruses : beacons in the biosphere / Anna Marie Skalka.
Description: Cambridge, Mas­sa­chu­setts : Harvard University Press, 2018. |
Includes bibliographical references and index.
Identifiers: LCCN 2018002504 | ISBN 9780674971707 (hardcover : alk. paper)
Subjects: LCSH: Retroviruses. | Retrovirus infections. | Viruses—­Evolution. |
Medicine—­Research—­History.
Classification: LCC QR414.5 S53 2018 | DDC 579.2/569—­dc23
LC rec­ord available at https://­lccn​.­loc​.­gov​/­2018002504

Jacket design: Tim Jones

Jacket image: Thinkstock (902636268)
This book was written for readers of all ages who
marvel at the won­ders of our biologic cosmos and
at the manner in which they have been revealed.

It is dedicated to my husband, Rudy; c­ hildren,

Jeannemarie and Christian; and grandchildren,
Kazimir and Shiloh, that they may glimpse the bea-
cons that have lit my way.
 — Con te n ts —

List of Tables and Figures ix

Introduction 1

1 Early Pioneers 3
2 Amending the Central Dogma 24
3 The Origin of Retroviruses 46
4 Retroviruses and Evolution 72
5 Revealing the Ge­ne­tic Basis of Cancer 96
6 HIV and the AIDS Pandemic 119

Epilogue 148

Notes 161
Suggested Readings 165
Acknowl­edgments 171
Index 173
 — Tabl e s an d Figures —

Tables
1.1 Selected Timelines for Discoveries in Genetics
and Retrovirology 4
2.1 The Retrovirus Family 44
3.1 Major Classes of Transposable Elements in the
Human Genome 61

Figures
1.1 Mendel’s crosses depicted in a Punnett square 8
1.2 Genes are encoded in DNA 11
1.3 The Watson-Crick double helix is held together
by nucleotide base pairing 15
1.4 Semi-conservative duplication of DNA 17
1.5 Organization of the ribosome 17
1.6 Transcription of messenger RNA from genes
in DNA 19
1.7 The genetic code 21
1.8 Translation of messenger RNA (mRNA) into
protein 22
2.1 Serial dilutions of a virus preparation and the
bacteriophage plaque assay 27
2.2 Plate assays for animal viruses 30
2.3 Retroviral DNA synthesis and its integration into
host DNA 37
2.4 Particle architecture and genome organization
of the prototype avian leukosis virus 40
 Tables an d Fig ure s

2.5 Reproduction cycle of the prototype avian

leukosis virus in a host cell 42
3.1 The Miller-Urey apparatus 48
3.2 Self-splicing by the Tetrahymena ribozyme 53
3.3 Successive steps in the transition from an RNA
world to a DNA world 55
3.4 The tree of life 56
3.5 The retrotransposons 63
3.6 Propagation of L1 LINE retrotransposons 65
3.7 Telomeres are formed by addition of repeated
sequences to chromosome ends via reverse
transcription 70
4.1 Nucleic acid hybridization 75
4.2 Estimating the age of endogenous retroviruses
from an evolutionary tree 76
4.3 Endogenous retroviral DNA recombination
induces host gene shuffling 79
4.4 Regulatory sequences in the LTRs of endogenous
retroviruses can affect host gene expression 81
4.5 Splicing signals in HERVs can lead to altered cell
messenger RNA production and abnormal
protein formation 81
4.6 The host-virus evolutionary arms race 92
5.1 Isolation of a src probe by subtractive
hybridization 100
5.2 Signal transduction 103
5.3 Origin of transducing retroviruses 107
5.4 Major lymphocytes of the adaptive immune
system 110
5.5 HTLV-I provirus and genes encoding viral
oncoproteins 114
6.1 HIV antibody assay provides the first blood
screen 127
6.2 Cradle of the AIDS pandemic 131

x
 Tables an d Fig ure s

6.3 HIV-1 origins via cross-species transmissions

of SIV 136
6.4 Regulatory and auxiliary proteins encoded in
the HIV-1 genome 140
6.5 Life-sparing effect of HIV antiviral treatment 143

xi
DISCOVERING RETROVIRUSES
 —­ In trodu c tion —

Retroviruses are ancient invaders of their host’s genomes. This virus

­family is unique among animal viruses in its capacity to copy its ge­ne­tic
blueprint of RNA into DNA (the hereditary material in all animals) and
then to splice that copy into the DNA of its host cell. While the host
cell normally survives this invasion, the retroviral DNA remains as a
permanent ge­ne­tic parasite, co-­opting cellular machinery to maintain
its existence and, in most cases, to produce new retrovirus particles that
can infect other cells. About 8 ­percent of the DNA in our bodies com-
prises copies of ancient retroviral DNA and bits thereof, whereas only
1.5 ­percent accounts for information that encodes all of the proteins
that make up our cells and ensure their function. This dichotomy came as
quite a shock to the scientists who sequenced the first h
­ uman genome—
a somewhat humbling realization that retroviruses “R” us.
But we are not alone; retroviral DNA or that of its pre­de­ces­sors is
pres­ent in all branches of the tree of life. In the course of evolution,
multicellular organisms have acquired numerous mechanisms to defend
against retroviral invasion—in an ongoing host versus virus “arms
race.” Nevertheless, in some notable cases, acquisition of retroviral
genes has actually afforded an opportunity for its host. Reprogramming
of retroviral genes in host DNA launched the evolution of placental
mammals, to which we ­humans belong.
Retroviruses are often, but not always, benign modern travelers. Ret-
roviruses can cause a variety of cancers in h ­ umans and other animals.
Analy­sis of the mechanisms by which they do so has revealed the
ge­ne­tic basis of this disease. On the other hand, the retroviral, HIV-­
induced AIDS has killed more p­ eople worldwide than any other disease
 In troduction

of viral origin. While t­ here have been tremendous strides in the delin-
eation of protective mea­sures, and ever-­improving treatments are
available, HIV / AIDS is still an incurable disease. It may therefore be
surprising to learn that genet­ically gutted versions of HIV and its rela-
tives are being employed as vehicles for gene therapy and to better
understand and treat AIDS and other diseases. H­ uman beings have found
ways to advance both science and medicine by harnessing even this hor-
rific killer.
The pages that follow describe the amazing stories of the discovery
of ­these ancient invaders / modern travelers and the fascinating “yin
and yang” of their existence. They ­will also disclose how studies of ret-
roviruses and concomitant discoveries in ge­ne­tics have illuminated the
landscape of our biological world. Traveling back in time to the origin
of life on Earth, through evolution and to the pres­ent day, we w
­ ill track
the many astonishing revelations garnered through pursuit of ­these
unique Beacons in the Biosphere.

2
 —­1 —

Early Pioneers

By the end of the nineteenth ­century, tiny organisms invisible to the

naked eye, such as bacteria, had already been seen and studied with light
microscopes. Furthermore, a connection between bacteria and disease
had been established by the German physician Robert Koch in his
studies of anthrax, a common disease of ­cattle at the time. Viruses,
however, w ­ ere mysterious entities and an entirely new kind of disease
agent. The first of ­these to be described, in 1892, was the plant virus
that ­causes tobacco mosaic disease. The second, discovered four years
­later, induces foot-­and-­mouth disease in ­cattle. Unlike bacteria, t­hese
plant and animal pathogens could not be seen in the existing micro-
scopes and, indeed, w ­ ere so small that they could pass through filters
known to retain bacteria and other microbes. Most impor­tant, ­these
agents ­couldn’t be propagated in broth solutions like bacteria; their
numbers increased only within the cells of their host organism. ­Because
­these new “filterable” agents seemed like some sort of contagious liquid
poison to early researchers, they w ­ ere given the Latin name for poison:
viruses.
The very first retrovirus discovered is actually related to our modern
scourge, the AIDS virus, HIV (­human immunodeficiency virus). While
studying an anemia in h ­ orses called swamp fever in the early 1900s, two
French veterinarians, Henri Vallée and Henri Carré, found that a filter-
able agent now known as the equine infectious anemia virus (EIAV)
could pass the disease from one animal to another. Four years l­ ater, two
Danish scientists, Vilhelm Ellerman and Olaf Bang, described another
filterable agent, one that could transmit leukemia among chickens. But
­these two early accounts of what would eventually be identified as ret-
roviral infections made l­ittle impression on the medical community. A
disease of h­ orses seemed irrelevant to h ­ umans, and leukemias ­were not
recognized as cancer at the time. This all changed when an American
 Early P ion eer s

­Table 1.1 ​Selected Timelines for Discoveries in Ge­ne­tics and Retrovirology

1866 Gregor Mendel reports results of studies of inheritance in pea plants.
1869 Friedrich Miescher identifies DNA (first called “nuclein”) in cell nuclei.
1882 Walther Flemming reports visualization of chromosome doubling and movement during
cell division.
1900 Mendel’s experiments are “rediscovered” by three botanists: a Dutchman, Hugo de Vries;
a German, Carl Correns; and an Austrian, Erich Tschermak.
1902–1903 Theodor Boveri and Walter Sutton propose the chromosome theory of heredity,
which conforms to Mendelian princi­ples.
1904 French veterinarians Henri Vallée and Henri Carré discover a filterable agent that transmits
anemia in ­horses, now classified as a retrovirus, equine infectious anemia virus (EIAV).
1905 The British scientist, William Bateson, translates Mendel’s paper into En­glish and names
the field of study “ge­ne­tics.”
1908 Danish scientists Vilhelm Ellerman and Olaf Bang discover an infectious filterable agent
that ­causes leukemia in chickens, ­later identified as the retrovirus avian leukemia virus (ALV).
1910–1911 T. H. Morgan and colleagues confirm the chromosome theory of heredity in studies
of sex-­linked inheritance of eye color in fruit flies.
1911 Peyton Rous discovers a tumor-­inducing infectious agent in chickens. The agent was ­later
known as the Rous sarcoma virus and assigned to the avian sarcoma virus (ASV) group of
retroviruses.
1913 Alfred Sturtevant (a student working with T. H. Morgan) constructs the first gene linkage
map, showing that genes are arranged like beads on a string in chromosomes.
1914 Rous, as well as Fujinami and Inamoto, in­de­pen­dently report additional virus-­induced
tumors. The Japa­nese isolate is ­later known as Fujinami sarcoma virus (FSV).
1914–1918 World War I
1928 British bacteriologist Fred Griffith reports that some component from heat-­killed virulent
pneumococcal bacteria can “transform” a nonvirulent strain into a virulent one.
1931 Harriet Creighton and Barbara McClintock show that ge­ne­tic recombination is associated
with the physical exchange of chromosomal segments.
1936 John Joseph Bittner discovers a “milk ­factor” that is transmitted by cancerous ­mothers to
young mice while nursing, ­later identified as the retrovirus mouse mammary tumor virus
(MMTV).
1939–1945 World War II
1944 Oswald Avery, William M ­ acLeod, and Maclyn McCarty report that DNA from virulent
strains mediates transformation of nonvirulent strains of Pneumococcus.
1950 Erwin Chargaff reports that DNA contains equal amounts of the bases A and T and of C
and G, although the ratios of A+T to C+G ratio can differ between organisms.
1951–1957 Ludwik Gross, at the veterans hospital in New York City, isolates a potent virus that
­causes mouse leukemia, mouse leukemia virus (MLV), l­ater assigned to the retrovirus f­amily.
1952 Alfred Hershey and Martha Chase confirm that DNA mediates heredity in bacteriophage
labeling experiments.
1953 James Watson and Francis Crick report their model of the DNA double-­helix structure.

4
 Early P ion eer s

­Table 1.1 ​(continued)
1954 George Palade observed ribosomes in the cytoplasm of cells by using an electron
microscope. ­These dense particles ­were ­later shown to be protein synthesis factories.
1954 Bjorn Sigurdsson describes an infectious agent that c­ auses a slowly developing but fatal
disease in sheep in Iceland. Called Visna virus, it is l­ater shown to be related to the AIDS
retrovirus, HIV.
1956 Arthur Kornberg and colleagues isolate an enzyme from the bacterium, Escherichia coli,
that copies a template DNA strand.
1957 Charlotte Friend reports that cell-­free transmission of leukemia in mice is caused by a
virus to be known as Friend murine leukemia virus (Fr-­MLV). This was one of many new
retroviruses to be discovered in the following years.
1958 Matthew Meselson and Franklin Stahl DNA show that DNA replication is semi-­conservative,
using a technique they in­ven­ted called “equilibrium density gradient centrifugation,” which
relied on the use of “heavy” isotopes.
1958 Paul C. Zamecnik and Mahlon Hoagland discover “adapter” RNAs with bound single amino
acids, ­later called transfer RNAs (tRNAs).
1960 Sidney Brenner, Francis Crick, François Jacob, and Jacques Monod predict that messenger
RNA (mRNA) is the intermediate between DNA and protein.
1961 The ge­ne­tic code is cracked by Marshall Nirenberg, together with colleagues Matthaei and
Leder, and by Har Gobind Khorana.
Note: Bold dates identify discoveries that w
­ ere recognized by Nobel Prizes.

pathologist, Francis Peyton Rous, made the startling announcement

that a filterable agent could cause solid tumors.
Peyton Rous was trained as a physician at Johns Hopkins Medical
School in Baltimore but deci­ded ­after his internship that taking care of
patients was neither his forte nor calling. Feeling “unfit” for clinical
medicine, he obtained an assistantship in pathology at the University
of Michigan, where he could focus instead on laboratory research. The
story of his discovery of a virus that could cause solid tumors began in
1909 when, only four years ­after receiving his medical degree, he was
invited to set up a cancer research laboratory at the Rocke­fel­ler Insti-
tute for Medical Research in New York (now Rocke­fel­ler University).
Shortly ­after establishing his new laboratory, Rous was visited by a
­woman from a nearby poultry farm who brought a hen with a large
lump in its breast for examination. Rous recognized the lump as a sar-
coma (a tumor of connective tissue), and perhaps b­ ecause he was new
to the profession with no baggage of biases, this hen sparked his in-
terest. Rous was aware of earlier reports that cancers can sometimes

5
 Early P ion eer s

spread among animals, much like an infection, and he deci­ded to test

this idea. He first observed that new sarcomas w ­ ere formed when he
placed bits of the tumor in the bodies of healthy chickens from
the same inbred stock. To find out if such spread was due to a virus,
he mixed ground bits of the tumor in a saline solution and then passed
the solution through a filter that would exclude bacteria. When new
tumors arose in chickens injected with the filtered solution, Rous con-
cluded that the infectious agent must certainly be a virus and reported
his findings in two papers in 1910 and 1911.1
As is often the case with results that are unexpected or disrupt cur-
rent paradigms, Rous’s work generated considerable controversy. Some
of his contemporaries argued that the filtrates ­were prob­ably not cell
­free or that the sarcomas w ­ ere not r­ eally cancer. O
­ thers considered
chickens to be too distantly related to provide a useful model for ­human
cancer. Indeed, the very idea that one could “catch” cancer from such
a tiny agent was met with general skepticism. Discouraged by the re-
sponse of his peers, Rous abandoned this line of research for de­cades,
focusing instead on World War I–­related proj­ects. His war­time work
was also quite productive; Rous developed the blood-­preserving tech-
niques that led to establishment of the world’s first blood bank near
the front line in Belgium. ­After the war, he returned to study cancer
caused by another virus, the Shope papilloma virus of rabbits. But, as
we ­will see in ­later chapters, discovery of the retrovirus named a­ fter
him, the Rous sarcoma virus (RSV), was to be considered his crowning
achievement, as it laid the foundation for a broad understanding of both
retroviruses and cancer. In 1966, Rous was awarded the Nobel Prize for
this work—­three years before his death at the age of ninety.
Although reports of similar cancer-­causing agents of mice and other
animal species occurred sporadically in the years following Rous’s
initial discovery, it would be de­cades ­until the unique properties of
retroviruses ­were appreciated. Such recognition was to await the de-
velopment of experimental tools and insights from research with vi-
ruses that infect bacteria, called bacteriophages (or simply phages; from
the Greek word to eat, “phagos”), and the establishment of the new
fields of ge­ne­tics and molecular biology.

6
 Early P ion eer s

To fully understand the impact of retroviruses on the fields of ge­

ne­tics, evolution, and medicine, we must look at the history of research
into the molecular basis of heredity, a story unfolding at the very same
time that the first retroviruses ­were being discovered.

Genes, DNA, and Establishment of the “Central Dogma”

In t­oday’s world, almost every­body knows about genes and DNA, if

not from news reports, then certainly from popu­lar tele­vi­sion shows,
where a murderer’s identity can be determined from a small, dried drop
of blood found at the crime scene. For this reason, it may be hard to
imagine that it was not known ­until the ­middle of the past ­century that
heritable traits (genes) are encoded in DNA. The fascinating path to this
knowledge was somewhat circuitous and pro­g ress extremely slow. In-
deed, the first critical observations ­were made in the ­middle of the
1800s by Gregor Mendel, an Austrian monk and scientist, working in
the experimental garden of an Augustinian abbey in what is now the
Czech Republic.
Mendel chose pea plants as an experimental system to study heredity
­because many distinct va­ri­e­ties ­were available, and offspring could be
produced quickly and easily in the abbey garden. He was interested in
knowing how par­tic­u­lar traits, such as seed color or shape, w­ ere passed
on through generations. Over a period of eight years, while performing
meticulous crosses and studying almost 30,000 plants, Mendel was able
to establish several fundamental princi­ples. First, he demonstrated that
colors and shapes w ­ ere inherited in­de­pen­dently of one another, as if
they w ­ ere specified by separate “units.” He also determined that when
purebred plants ­were crossed, the property of one parent (e.g., a par­
tic­u­lar color) could be dominant in the next generation of hybrid plants.
Results from his numerous crosses between t­ hese hybrid plants showed
Mendel that the property that seemed to dis­appear in the hybrids, called
the “recessive” trait, would reappear among their progeny—­and, im-
portantly, in par­tic­u­lar ratios. On average, one plant exhibited the re-
cessive property for ­every three with the dominant property (1:3). By

7
 Early P ion eer s

Hybrid parent 1

R r

r rR rr

Hybrid parent 2

R RR Rr

Fig. 1.1 ​Mendel’s crosses depicted in a Punnett square. The Punnett square, named a­ fter the
ge­ne­ticist Reginald Crundall Punnett (1875–1967), is used by biologists to determine the prob-
ability that an offspring w­ ill exhibit a par­tic­u­lar trait as shown in the boxes. Mendel deduced
that ge­ne­tic traits must exist in pairs, one from each parent. When one of a pair is dominant
over the other (e.g., green over yellow), inheritance from a cross of hybrid parents can be de-
scribed by the ratio of 1:2:1 of pure recessive (rr), hybrid (rR or Rr), and pure dominant (RR). As
R is dominant over r, the ratio of green to yellow progeny produced by the hybrid parents is 3:1.

applying statistical methods and mathematical models, most unusual

for a botanist at the time, Mendel deduced that the relationship between
the dominant and recessive properties could be explained if two in­de­
pen­dent trait-­defining units existed in each parent, e­ ither one of which
was passed on to the next generation. Inheritance could be described
by the ratio of 1:2:1 of recessive, hybrid, and dominant, as illustrated
in the boxes of a Punnett square. This is a phenomenon with which
we are now familiar in ­human eye color; that is, two parents with brown
eyes (the dominant trait) can produce a child with blue eyes if they both
carry and pass on their recessive gene for blue eyes.
A somewhat modest and deferential scientist, Mendel was discour-
aged when a leading authority of the time, the Swiss botanist Karl Will-
helm von Nägeli, rejected Mendel’s results and conclusions. Nägeli
had his own ideas for how ge­ne­tic traits are inherited (now known to
be incorrect), and he was writing a book on this topic. He ­didn’t be-
lieve Mendel’s results to be generally applicable ­because, as Mendel ­later
confirmed, the plant Nägeli was studying did not transmit ge­ne­tic traits
in the same way as Mendel’s peas. What neither scientist knew at the
time was that this difference was due to the fact that Nägeli’s plants re-
produce mainly via a mechanism in which the seeds carry ge­ne­tic
traits of only one parent. Mendel eventually delivered two lectures on

8
 Early P ion eer s

his findings at the 1865 meeting of the Natu­ral Science Society in Brno,
and his results ­were published in its journal the following year. Never-
theless, although he is now universally recognized as the founder of
ge­ne­tics, Mendel’s work was overlooked for the next forty years. The
princi­ples established by his experiments, now known as Mendel’s laws
of inheritance (“in­de­pen­dent assortment,” “segregation,” and “domi-
nance”), ­were rediscovered in 1900 (sixteen years a­ fter Mendel died)
when other researchers, using peas and other plants, sought to explain
their similar findings. As happens not infrequently in science, discov-
eries that seem odd or even irrelevant at the time they are made can
provide essential pieces to impor­tant puzzles that arise de­cades ­later.
This posthumous appreciation of Mendel’s work inspired a greatly re-
newed interest in ge­ne­tics, but it would take another fifty years u ­ ntil
we would know what genes are made of.
By the late 1800s, use of the light microscope allowed the visualiza-
tion not only of some microorganisms but also of features inside of
cells. Among ­these ­were linear structures within the nuclei, called chro-
mosomes (from the Greek words for colored, “chroma,” and bodies,
“soma”), which could be seen most clearly when cells ­were in the
pro­cess of dividing. The realization that genes reside within t­hese
linear chromosomes came from in­de­pen­dent studies in 1902–1903 by
Walter Sutton, a gradu­ate student at Columbia University, and an es-
tablished German scientist, Theodor Boveri. Sutton based his theory
on observations of insect chromosomes, which are lined up in matched
pairs that separate during formation of eggs and sperm, precisely as
expected by Mendel’s laws. Boveri’s studies showed that a proper com-
plement of chromosomes had to be pres­ent if the sea urchin embryos
he was studying w ­ ere to develop normally. The chromosome theory
of inheritance gained further support in studies of fruit fly eye color
by Thomas Hunt Morgan and was accepted broadly in 1931 when Har-
riet Creighton and Barbara McClintock showed that ge­ne­tic recombi-
nation is correlated with physical exchanges between chromosomes in
dividing corn cells.
Chromosomes ­were known to contain both proteins and DNA, but
it was the protein component that was assumed to comprise genes in
the 1930s. Consisting of long chains with dif­fer­ent combinations of

9
 Early P ion eer s

twenty amino acids, proteins appeared to be the only cellular constitu-

ents sufficiently complex to embody hereditary traits. DNA, made up
of long, repetitious chains of only four dif­fer­ent nucleotides (sugar-­
phosphate molecules, which contain one of four bases), was consid-
ered a most unlikely candidate. Consequently, the best guess at the time
was that DNA was some sort of general “scaffold” that supported genes.
The first evidence that DNA is heredity material was obtained in
1944 by Oswald Avery and his Rocke­fel­ler University colleagues, Colin
­MacLeod and Maclyn McCarty. Avery became intrigued by a 1928 re-
port that a component from killed pathogenic pneumococcal bacteria
could induce virulence in a benign strain of the same bacteria when
the two w­ ere mixed together, a pro­cess called “transformation.” He and
his colleagues deci­ded to track down the responsible component by a
pro­cess of elimination. They prepared extracts from the killed virulent
strain and treated the extracts with enzymes that would destroy pro-
teins, lipids, or DNA. They then exposed the benign bacteria to the
extracts to discover which treatments abolished transformation, which
they monitored by the formation of large, smooth colonies of virulent
bacteria. Their experiments showed that transformation activity was
lost only a­ fter treatment with enzymes able to destroy DNA. Further-
more, addition of chemically purified DNA from the virulent strain
caused transformation of cultured benign cells. From ­these results,
Avery and his colleagues concluded that genes specifying virulence
must be encoded in DNA. As one may suspect, their seemingly
compelling results w ­ ere met not with acclaim but with general disbe-
lief. Critics grumbled that Avery’s DNA preparations might be con-
taminated with sufficient amounts of protein to cause the resulting
transformation. A Rocke­fel­ler colleague, Alfred Mirsky, copublished a
widely read, critical article in 1946 noting that “as much as 1 or
2 ­percent of protein could be pres­ent in a preparation of ‘pure, protein-­
free’ nucleic acid.”2 Indeed, the idea that genes ­were embodied in pro-
teins was so thoroughly accepted at the time, many leading biologists
simply disregarded Avery’s work.
The situation changed in 1952 with a publication from Alfred Her-
shey’s laboratory at the Car­ne­g ie Institution’s Department of Ge­ne­tics

10
 Early P ion eer s

A
DNA DNA
from from
S cells S cells
+
DNase
Bacteria R R R

R R+S R

B Infection Blending Separation Analysis of new

phage particles

Viral protein contains Radioactivity No radioactivity

radioactive sulfur remains in the in progeny phage
supernatant fraction

Viral DNA contains Radioactivity found Radioactive phosphorus

radioactive phosphorus in the cell pellet incorporated into
progeny phage

Fig. 1.2 ​Genes are encoded in DNA. (A) An illustration of the 1944 experiment of Avery, McLeod,
and McCarty. The nonvirulent pneumococcal bacteria suspended in the first test tube make small
rough colonies when planted in culture dishes (R). A mixture of small rough and large smooth
(S) colonies is formed if DNA purified from the virulent strain is added to the nonvirulent bacteria,
shown in the center. As illustrated on the right, only rough colonies are formed if the purified
DNA from the virulent strain is first treated with an enzyme (DNase) known to degrade DNA.
(B) An illustration of the Hershey-­Chase blender experiment. Radioactive protein in an infecting
phage particle is shown as an orange head, and radioactive DNA injected from an infecting particle
is red. Only the DNA enters infected cells and is incorporated into progeny phage particles.

11
 Early P ion eer s

in Cold Spring Harbor. Hershey was a founding member of a tight-­knit

cohort of scientists called the Phage Group. It had been known since
1915 that bacterial cells are also hosts to viruses (bacteriophages). Mem-
bers of the Phage Group ­were convinced that bacteriophages offered
the best chance at understanding the molecular basis of heredity
­because they likely had small, manageable sets of genes. The host bac-
terium of choice for the Phage Group was Escherichia coli (E. coli), a
common constituent of the ­human gut and easy to culture in the lab-
oratory. E. coli was to become a work­horse for the field and is t­ oday
one of the best-­understood organisms in the world. The E. coli phage
that Hershey studied, called T2, was known to contain DNA sur-
rounded by a protein coat.
Although aware of Avery’s work, Hershey still believed that genes
­were encoded in protein, as did most scientists at the time. To test this
notion, he exploited the newly available radioactive forms of phos-
phorus (32P) and sulfur (35S). Hershey and his research assistant Martha
Chase prepared radioactive phage particles by infecting bacteria in me-
dium that contained ­either isotope. ­Because DNA is rich in phosphate
but not sulfur, and the opposite is true of proteins, ­these phage parti-
cles contained “labels” that ­were specific for each type of molecule. The
radioactive phage particles ­were then used to infect fresh bacteria, and
­after the particles w­ ere allowed to attach to their host cells, the mix-
ture was subjected to vigorous agitation using an ordinary kitchen item,
called a Waring blender. The infected bacteria w ­ ere then isolated from
the blended mixture to determine where the labeled phage DNA and
protein had gone. What Hershey and Chase found was both indisput-
able and, to them, quite astonishing. Most of the 35S-­labeled phage
protein was shaken from the infected bacteria by blending. In con-
trast, almost all of the 32P-­labeled phage DNA was associated with the
isolated bacteria. Furthermore, progeny phage particles produced by
t­ hese bacteria contained 32P, as would be expected for ge­ne­tic inheri-
tance via DNA. T ­ hese results w­ ere exactly the opposite of what Her-
shey expected; he thought that the DNA “scaffold,” would be washed
away in the blender and much of the protein would be retained in the
bacteria. Typically cautious in his interpretation, Hershey concluded

12
 Early P ion eer s

that “the protein component(s) prob­ably has no function in reproduc-

tion of the phage” but “the DNA does.”3 Nevertheless, this conceptu-
ally ­simple but elegant study converted all previous skeptics, and ­today,
the Hershey-­Chase experiment is among the most famous in scientific
history. ­Later studies with a variety of phages and also some animal
viruses showed that introduction of purified viral DNA into a host
cell is all that is required for production of a new crop of progeny
viruses.
Results from the “blender experiments” reached James (Jim) Watson,
in a letter from Hershey in 1951. A precocious young member of the
Phage Group, Watson had recently arrived in E ­ ngland as a postdoctoral
fellow in the Cavendish Laboratory at Cambridge University, where he
shared an office with Francis Crick. Watson had been interested in DNA
as the source of genes for some time, and the news from Hershey fu-
eled his growing passion to solve its structure. He was driven by the
conviction that such knowledge would provide a key to the “secret of
life.” Believed by many to be “too bright to be ­really sound,”4 the
twenty-­three-­year-­old Watson has been described most vividly by the
eminent French biologist, François Jacob, as “an amazing character. A
surprising mixture of awkwardness and shrewdness . . . ​childishness in
the ­things of life and maturity in the ­things of science.”5
Watson’s thirty-­five-­year-­old lab-­mate, Crick, was an equally note-
worthy character. He was described by Watson as having a quick, pen-
etrating mind but never in “a modest mood” and “most ­people thought
he talked too much.”6 Crick had been trained in mathe­matics and
physics but was still working on his PhD research, which dealt not with
DNA but with protein structure determination. Brash and unknown
at the time, yet enticed by this most exciting challenge, Watson and
Crick set out together to solve the structure of DNA.
The story of their success, as well as its momentous implications,
has been told in numerous publications, including separate books
authored by Watson and by Crick and even in TV movies. The two
believed that that they could get the structure without d­ oing any ex-
periments by building a model of DNA from its known constituents,
­because the famous Caltech scientist Linus Pauling had recently been

13
 Early P ion eer s

successful in solving protein structures this way. They ­were also urged
on by the knowledge that Pauling was already working on models of
DNA, and they did not want to be scooped by this established presence
in the field. ­After a few false starts, their efforts w
­ ere galvanized when
Watson was shown the exquisite experimental data of Rosalind Franklin
(although without her knowledge) by Maurice Wilkins, head of a group
at King’s College. Franklin’s data, which aimed to derive DNA struc-
ture using X-­ray crystallography, revealed DNA as a helical molecule
made up of two strands. To construct their final model, Watson and
Crick drew upon this insight, as well as an impor­tant correlation
established by the biochemist, Erwin Chargaff, namely that the com-
ponent four bases exist in complementary pairs in DNA: the amount
of adenine (A) is always equal to the amount of thymine (T), and the
amount of guanine (G) is always equal to the amount of cytosine (C).
Their final model, built with brass plates and clamps, was a double helix
formed by phosphate-­linked sugar chains ­running in opposite directions
and held together by chemical bonds between the complementary base
pairs, much like the rungs on a spiral staircase.
In true British tradition of fair play, Wilkins was promptly invited
to Cambridge to inspect the Watson-­Crick model. He was immediately
convinced of its credibility and excited by the implications. Two days
­later, Wilkins informed Watson and Crick that he and Franklin con-
firmed that their X-­ray data strongly supported the double-­helix struc-
ture. They w
­ ere therefore anxious to publish the results of their physical
analyses together with a report of the model. Relieved at this gen-
erous response, Watson and Crick readily agreed. Two papers from the
King’s College group ­were subsequently published together with the
description of the Watson-­Crick DNA model in the April 25, 1953,
issue of the journal Nature.
The Watson-­Crick DNA structure turned out to be enormously in-
formative. The complementarity of bases in the two strands suggested
that DNA could be duplicated via the separation and copying of each
strand, thereby allowing ge­ne­tic information to be passed on as cells
divided. Furthermore, a pos­si­ble alphabet comprising the four DNA
bases could now be ­imagined, with “words” specifying par­tic­u­lar amino

14
Another random document with
no related content on Scribd:
been so much approved, that we can recommend it with some
confidence, as it stands. Modern taste would perhaps be rather in
favour of rich brown gravy and thick tomata sauce, or sauce
poivrade.
82. A deep oblong dish of suitable size seems better adapted to this purpose.
In dishing the pig lay the body flat in the middle, and the head and
ears at the ends and sides. When very pure oil can be obtained, it is
preferable to butter for the basting: it should be laid on with a bunch
of feathers. A pig of three weeks old is considered as best suited to
the table, and it should always be dressed if possible the day it is
killed.
1-1/4 to 1-3/4 hour.
 BAKED PIG.

Prepare the pig exactly as for roasting; truss, and place it in the
dish in which it is to be sent to the oven, and anoint it thickly in every
part with white of egg which has been slightly beaten; it will require
no basting, nor further attention of any kind, and will be well crisped
by this process.
 PIG À LA TARTARE

When the shoulders of a cold roast pig are left entire, take them off
with care, remove the skin, trim them into good form, dip them into
clarified butter or very pure salad oil, then into fine crumbs highly
seasoned with cayenne and mixed with about a half-teaspoonful of
salt. Broil them over a clear brisk fire, and send them quickly to table,
as soon as they are heated through and equally browned, with
tomata sauce, or sauce Robert. Curried crumbs and a currie-sauce
will give an excellent variety of this dish; and savoury herbs with two
or three eschalots chopped small together, and mixed with the
bread-crumbs, and brown eschalot sauce to accompany the broil,
will likewise be an acceptable one to many tastes.
 SUCKING PIG EN BLANQUETTE. (ENTRÉE.)

Raise the flesh from the bones of a cold roast pig, free it from the
crisp outer skin or crackling, and cut it down into small handsome
slices. Dissolve a bit of butter the size of an egg, and throw in a
handful of button-mushrooms, cleaned and sliced; shake these over
the fire for three or four minutes, then stir to them a dessertspoonful
of flour and continue to shake or toss them gently, but do not allow
them to brown. Add a small bunch of parsley, a bay-leaf, a middling-
sized blade of mace, some salt, a small quantity of cayenne or white
pepper, half a pint of good veal or beef broth, and from two to three
glasses of light white wine. Let these boil gently until reduced nearly
one third; take out the parsley and mace, lay in the meat, and bring it
slowly to the point of simmering; stir to it the beaten yolks of three
fresh eggs, and the strained juice of half a lemon Serve the
blanquette very hot.
 TO ROAST PORK.

When the skin is left on the joint which is to be roasted, it must be

scored in narrow strips of equal width, before it is put to the fire, and
laid at a considerable distance from it at first, that the meat may be
heated through before the skin hardens or begins to brown; it must
never stand still for an instant, and the basting should be constant.
Pork is not at the present day much served at very good tables,
particularly in this form; and it is so still less with the old savoury
stuffing of sage and onions, though some eaters like it always with
the leg: when it is ordered for this joint, therefore prepare it as
directed for a goose, at page 160, and after having loosened the skin
from the knuckle, insert as much as can well be secured in it. A little
clarified butter or salad oil may be brushed over the skin quite at first,
particularly should the meat not be very fat, but unless remarkably
lean, it will speedily yield sufficient dripping to baste it with. Joints
from which the fat has been pared, will require of course far less
roasting than those on which the crackling is retained. Brown gravy,
and apple or tomata sauce, are the usual accompaniments to all
roasts of pork except a sucking pig; they should always be
thoroughly cooked.
Leg of pork of 8 lbs., 3 hours; loin of from 5 to 6 lbs., with the skin
on, 2 to 2-1/2 hours; spare rib of 6 to 7 lbs., 1-1/2 hour.
 TO ROAST A SADDLE OF PORK.

The skin of this joint may be removed entirely, but if left on it must
be scored lengthwise, or in the direction in which it will be carved.
The pork should be young, of fine quality, and of moderate size.
Roast it very carefully, either by the directions given in the preceding
receipt, or when the skin is taken off, by those for a saddle of mutton,
allowing in the latter case from three quarters of an hour to a full
hour more of the fire for it in proportion to its weight. Serve it with
good brown gravy and tomata sauce, or sauce Robert; or with apple
sauce should it be preferred. 20 minutes to the pound, quite [TN: text
missing.]
 TO BROIL OR FRY PORK CUTLETS.

Cut them about half an inch thick from a delicate loin of pork, trim
them into neat form, and take off part of the fat, or the whole of it
when it is not liked; dredge a little pepper or cayenne upon them,
and broil them over a clear and moderate fire from fifteen to eighteen
minutes: sprinkle a little fine salt upon them just before they are
dished. They may be dipped into egg and then into bread-crumbs
mixed with minced sage, and finished in the usual way.[83] When
fried, flour them well, and season them with salt and pepper first.
Serve them with gravy in the pan, or with sauce Robert.
83. If broiled, with the addition of these a little clarified butter must be added to
the egg, or sprinkled on the cutlets.
 COBBETT’S RECEIPT FOR CURING BACON.

“All other parts being taken away, the two sides that remain, and
which are called flitches, are to be cured for bacon. They are first
rubbed with salt on their inside, or flesh sides, then placed one on
the other, the flesh sides uppermost in a salting trough, which has a
gutter round its edges to drain away the brine; for to have sweet and
fine bacon, the flitches must not be sopping in brine, which gives it
the sort of taste that barrel-pork and sea-junk have, and than which
is nothing more villainous. Everyone knows how different is the taste
of fresh dry salt from that of salt in a dissolved state. Therefore,
change the salt often; once in four or five days. Let it melt and sink
in, but let it not lie too long. Change the flitches, put that at the
bottom which was first on the top. Do this a couple of times. This
mode will cost you a great deal more in salt than the sopping mode;
but without it your bacon will not be so sweet and fine, nor keep so
well. As to the time required for making the flitches sufficiently salt, it
depends on circumstances; the thickness of the flitch, the state of
the weather, the place wherein the salting is going on. It takes a
longer time for a thick than for a thin flitch; it takes longer in dry than
in damp weather, it takes longer in a dry than in a damp place. But
for the flitches of a hog of five score, in weather not very dry or very
damp, about six weeks may do; and as yours is to be fat, which
receives little injury from over-salting, give time enough; for you are
to have bacon till Christmas comes again. The place for salting
should, like a dairy, always be cool, but always admit of a free
circulation of air; confined air, though cool, will taint meat sooner
than the midday sun accompanied with a breeze. With regard to
smoking the bacon, two precautions are necessary: first to hang the
flitches where no rain comes down upon them, and next, that the
smoke must proceed from wood, not peat, turf, nor coal. As to the
time that it requires to smoke a flitch, it must depend a good deal
upon whether there be a constant fire beneath, and whether the fire
be large or small. A month will do if the fire be pretty constant, and
such as a farm house fire usually is. But oversmoking, or rather, too
long hanging in the air, makes the bacon rust. Great attention
should, therefore, be paid to this matter. The flitch ought not to be
dried up to the hardness of a board, and yet it ought to be perfectly
dry. Before you hang it up, lay it on the floor, scatter the flesh-side
pretty thickly over with bran or with some fine saw-dust, not of deal
or fir. Rub it on the flesh, or pat it well down upon it. This keeps the
smoke from getting into the little openings, and makes a sort of crust
to be dried on.
“To keep the bacon sweet and good, and free from hoppers, sift
fine some clean and dry wood-ashes. Put some at the bottom of a
box or chest long enough to hold a flitch of bacon. Lay in one flitch;
and then put in more ashes, then another flitch, and cover this with
six or eight inches of the ashes. The place where the box or chest is
kept ought to be dry, and should the ashes become damp they
should be put in the fire-place to dry, and when cold, put back again.
With these precautions the bacon will be as good at the end of the
year as on the first day.”
Obs.—Although the preceding directions for curing the bacon are
a little vague as regards the proportions of salt and pork, we think
those for its after-management will be acceptable to many of our
readers, as in our damp climate it is often a matter of great difficulty
to preserve hams and bacon through the year from rust.
 A GENUINE YORKSHIRE RECEIPT FOR CURING HAMS AND
BACON.

“Let the swine be put up to fast for twenty-four hours before they
are killed (and observe that neither a time of severe frost, nor very
damp weather, is favourable for curing bacon). After a pig has been
killed and scalded, let it hang twelve hours before it is cut up, then
for every stone or fourteen pounds’ weight of the meat, take one
pound of salt, an ounce and a quarter of saltpeter, and half an ounce
of coarse sugar. Rub the sugar and saltpetre first into the fleshy
parts of the pork, and remove carefully with a fork any extravasated
blood that may appear on it, together with the broken vessels
adjoining; apply the salt especially to those parts, as well as to the
shank-ends of the hams, and any other portions of the flesh that are
more particularly exposed. Before the salt is added to the meat,
warm it a little before the fire, and use only a part of it in the first
instance; then, as it dissolves, or is absorbed by the meat, add the
remainder at several different times. Let the meat in the meanwhile
lie either on clean straw, or on a cold brick or stone floor: it will
require from a fortnight to three weeks’ curing, according to the state
of the atmosphere. When done, hang it in a cool dry place, where
there is a thorough current of air, and let it remain there until it is
perfectly dry, when the salt will be found to have crystallized upon
the surface. The meat may then be removed to your store, and kept
in a close chest, surrounded with clean outer straw. If very large, the
hams will not be in perfection in less than twelve months from the
time of their being stored.”
Pork 20 stone; salt, 20 lbs.; saltpetre, 20 oz.; sugar, 10 oz.; 14 to
21 days.
 KENTISH MODE OF CUTTING UP AND CURING A PIG.

To a porker of sixteen stone Kentish weight (that is to say, eight

pounds to the stone, or nine stone two pounds of common weight),
allow two gallons of salt, two pounds of saltpetre, one pound of
coarse sugar, and two pounds of bay-salt well dried and reduced to
powder. Put aside the hams and cheeks to be cured by themselves;
let the feet, ears, tail, and eye-parts of the head be salted for
immediate eating; the blade-bones, and ends of the loins and ribs
reserved for sausage-meat should it be wanted, and the loin and
spare-ribs for roasting. Divide and salt the remainder thus: Mix well
together the saltpetre, sugar, and bay-salt, and rub the pork gently
with them in every part; cover the bottom of the pickling tub with salt,
and pack in the pork as closely as possible, with a portion of the
remaining salt between each layer. A very little water is sometimes
sprinkled in to facilitate the dissolving of the salt into a brine, but this
is always better avoided, and in damp weather will not be needed. If
in a fortnight it should not have risen, so as almost entirely to cover
the meat, boil a strong brine of salt, saltpetre, sugar, and bay-salt; let
it remain until perfectly cold, and then pour it over the pork. A board,
with a heavy stone weight upon it, should be kept upon the meat to
force it down under the brine. In from three to four months it will be fit
for table, and will be delicate and excellent pickled pork.
The pickling parts of a porker of sixteen stone (Kentish weight, or
nine stone two pounds of common weight, or fourteen pounds to the
stone); common salt, 2 gallons; saltpetre, 2 lbs.; coarse sugar, 1 lb.:
bay-salt, 2 lbs.
 FRENCH BACON FOR LARDING.

Cut the bacon from the pig with as little lean to it as possible. Rub
it well in every part with salt which has been dried, reduced to
powder, and sifted; put the layers of bacon close against and upon
each other, in a shallow wooden trough, and set in a cool, but not a
damp cellar; add more salt all round the bacon, and lay a board, with
a very heavy weight upon it. Let it remain for six weeks, then hang it
up in a dry and airy place.
Pork, 14 lbs.; salt, 14 oz.: 6 weeks.
 TO PICKLE CHEEKS OF BACON AND HAMS.

One pound of common salt, one pound of the coarsest sugar, and
one ounce of saltpetre, in fine powder, to each stone (fourteen
pounds) of the meat will answer this purpose extremely well. An
ounce of black pepper can be added, if liked, and when less sugar is
preferred, the proportion can be diminished one half, and the
quantity of salt as much increased. Bacon also may be cured by this
receipt, or by the Bordyke one for hams. A month is sufficient time
for the salting, unless the pork be very large, when five weeks must
be allowed for a ham. The ingredients should be well mixed, and all
applied at the same time.
To each 14 lbs. of pork, salt, 1 lb.; coarse sugar, 1 lb.; saltpetre, 1
oz.; pepper (if used), 1 oz.: 4 to 5 weeks.
 MONSIEUR UDE’s RECEIPT, HAMS SUPERIOR TO
WESTPHALIA.

(Excellent.)
“Take the hams as soon as the pig is sufficiently cold to be cut up,
rub them well with common salt, and leave them for three days to
drain; throw away the brine, and for a couple of hams of from fifteen
to eighteen pounds weight, mix together two ounces of saltpetre, a
pound of coarse sugar, and a pound of common salt; rub the hams in
every part with these, lay them into deep pickling-pans with the rind
downwards, and keep them for three days well covered with the salt
and sugar; then pour over them a bottle of good vinegar, and turn
them in the brine, and baste them with it daily for a month; drain
them well, rub them with bran, and let them be hung for a month high
in a chimney over a wood-fire to be smoked.”
Hams, of from 15 to 18 lbs. each, 2; to drain 3 days. Common salt,
and coarse sugar, each 1 lb.; saltpetre, 2 oz.: 3 days. Vinegar, 1
bottle: 1 month. To be smoked 1 month.
Obs.—Such of our readers as shall make trial of this admirable
receipt, will acknowledge, we doubt not, that the hams thus cured
are in reality superior to those of Westphalia. It was originally given
to the public by the celebrated French cook, Monsieur Ude. He
directs that the hams when smoked should be hung as high as
possible from the fire, that the fat may not be melted; a very
necessary precaution, as the mode of their being cured renders it
peculiarly liable to do so. This, indeed, is somewhat perceptible in
the cooking, which ought, therefore, to be conducted with especial
care. The hams should be very softly simmered,[84] and not over-
done. They should be large, and of finely-fed pork, or the receipt will
not answer. We give the result of our first trial of it, which was
perfectly successful, the ham cured by it being of the finest possible
flavour.
84. We have not had the trial made ourselves, but we think they would be even
finer baked than boiled.
 Leg of Suffolk farm-house pork, 14 to 15 lbs.; saltpetre, 1-1/4 oz.;
strong coarse salt, 6 oz.; coarse sugar, 8 oz.: 3 days. Fine whitewine
vinegar, 1 pint. In pickle, turned daily, 1 month. Smoked over wood, 1
month.
Obs.—“When two hams are pickled together, a smaller proportion
of the ingredients is required for each, than for one which is cured by
itself.”
 SUPER-EXCELLENT BACON.

For several successive years, after first testing the above receipt,
we had it adopted for curing bacon, with even more highly
satisfactory results, as it was of incomparable flavour, and remained
good for a great length of time, the vinegar preserving it entirely from
becoming rusted. Well-fed pork of delicate size was always used for
it, and excellent vinegar. The ingredients were added in the
proportions given in the receipt for the Suffolk ham which preceeds
this, and the same time was allowed for the salting and smoking.
 HAMS.

(Bordyke Receipt.)
After the hams have been rubbed with salt, and well drained from
the brine, according to our previous directions, take, for each
fourteen pounds weight of the pork, one ounce of saltpetre in fine
powder mixed with three ounces of very brown sugar; rub the meat
in every part with these, and let it remain some hours, then cover it
well with eight ounces of bay-salt, dried and pounded, and mixed
with four ounces of common salt: in four days add one pound of
treacle, and keep the hams turned daily, and well basted with the
pickle for a month. Hang them up to drain for a night, fold them in
brown paper, and send them to be smoked for a month. An ounce of
ground black pepper is often mixed with the saltpetre in this receipt,
and three ounces of bruised juniper-berries are rubbed on to the
meat before the salt is added, when hams of a very high flavour are
desired.
Ham, 14 lbs.; saltpetre, 1 oz.; coarse sugar, 3 oz.: 8 to 12 hours.
Bay-salt, 1/2 lb.; common salt, 4 oz.: 4 days. Treacle, 1 lb.: 1 month.
To heighten flavour, black pepper, 1 oz; juniper-berries, 3 oz.
 TO BOIL A HAM.

The degree of soaking which must be given to a ham before it is

boiled, must depend both on the manner in which it has been cured,
and on its age. If highly salted, hard, and old, a day and night, or
even longer, may be requisite to dilate the pores sufficiently, and to
extract a portion of the salt. To do either effectually the water must
be several times changed during the steeping. We generally find
hams cured by any of the receipts which we have given in this
chapter quite enough soaked in twelve hours; and they are more
frequently laid into water only early in the morning of the day on
which they are boiled. Those pickled by Monsieur Ude’s receipt need
much less steeping than any others. After the ham has been
scraped, or brushed, as clean as possible, pare away lightly any part
which, from being blackened or rusty, would disfigure it; though it is
better not to cut the flesh at all unless it be really requisite for the
good appearance of the joint. Lay it into a ham-kettle, or into any
other vessel of a similar form, and cover it plentifully with cold water;
bring it very slowly to boil, and clear off carefully the scum which will
be thrown up in great abundance. So soon as the water has been
cleared from this, draw back the pan quite to the edge of the stove,
that the ham may be simmered softly but steadily, until it is tender.
On no account allow it to boil fast. A bunch of herbs and three or four
carrots, thrown in directly after the water has been skimmed, will
improve it. When it can be probed very easily with a sharp skewer, or
larding-pin, lift it out, strip off the skin, and should there be an oven
at hand, set it in for a few minutes after having laid it on a drainer;
strew fine raspings over it, or grate a hard-toasted crust, or sift upon
it the prepared bread of Chapter V., unless it is to be glazed, when
neither of these must be used.
Small ham, 3-1/2 to 4 hours; moderate sized, 4 to 4-1/2 hours;
very large, 5 to 5-1/2 hours.
Obs.—We have seen the following manner of boiling a ham
recommended, but we have not tried it:—“Put into the water in which
it is to be boiled, a quart of old cider and a pint of vinegar, a large
bunch of sweet herbs, and a bay leaf. When it is two-thirds done,
skin, cover it with raspings, and set it in an oven until it is done
enough: it will prove incomparably superior to a ham boiled in the
usual way.”
 TO GARNISH AND ORNAMENT HAMS IN VARIOUS WAYS.

When a ham has been carefully and delicately boiled, the rind
while it is still warm, may be carved in various fanciful shapes to
decorate it; and a portion of it left round the knuckle in a semi-
circular form of four or five inches deep, may at all times be easily
scollopped at the edge or cut into points (vandykes). This, while
preserving a character of complete simplicity for the dish, will give it
an air of neatness and finish at a slight cost of time and trouble. A
paper frill should be placed round the bone.
The Germans cut the ham-rind after it has been stripped from the
joint, into small leaves and similar “prettinesses,”[85] and arrange
them in a garland, or other approved device, upon its surface. In
Ireland and elsewhere, bread evenly sliced, and stamped out with
cutters much smaller than a fourpenny-piece, then carefully fried or
coloured in the oven, is used to form designs upon hams after they
are glazed. Large dice of clear firm savoury jelly form their most
appropriate garnish, because they are intended to be eaten with
them. For the manner of making this, and glaze also see Chapter IV.
85. This should be done with a confectionary or paste cutter.
The ham shown in Plate V., which follows the directions for
“Carving,” is of very good appearance; but in common English
kitchens generally, even the degree of artistic skill required to form
its decorations well, is not often to be met with.