MOLECULAR GENETICS

The molecule called deoxyribonucleic acid (DNA) was discovered by Friedrich miescher in

1869. In 1868 Miescher demonstrated that the nuclear material could be isolated from the

cytoplasm. This nuclear material was later shown to be composed of protein and nucleic acids.

Because of the repetitive nature of nucleic acids and the apparent similarity in chemical structure

irrespective of source, it was thought not to play an important role in heredity. Amino acids were

thought to specify genetic information.

Transformation

Transformation is the evidence to identify the DNA as the genetic material or uptake of foreign

piece of DNA by an organism occasioning the alteration of genotype. In Diplococcus

pneumoniae bacteria the thin capsule enclosing the cell wall consists of substances called

polysaccharides, which are of specific types. Type II capsules for example elicit the formation of

antibodies in the bloodstream of rabbits that are different from antibodies formed with type III

capsules. These capsules types are distinct properties of bacterial strains and bacteria of one kind

do not appear in pure cultures of another kind.

On the other hand the capsules themselves are subject to variability in respect to their

presence or absence. That is, any particular type of D. pneumoniae may occasionally give rise to

bacteria that do not have capsule at all (mutation) such non-capsulated bacteria have rough (R)

appearance when grown on culture plates and relatively harmless in contrast to the smooth (S)

appearance and virulence of encapsulated bacteria. When cultured separately, R and S colonies

will transmit their respective characters to future generations except for rare mutations. These

bacteria are sensitive to heat and if temperature is raised sufficiently high the bacteria are heat-

killed and can no longer divide.

Griffith, in 1928, showed that heat-killed bacteria of one type could have a hereditary influence

on bacteria of another type. In one experiment he injected heat killed bacteria of type IIIS into a

mouse that carry type IIR non heat killed and obtained virulent live cultures which were of the

type IIIS variety. Because the injection of heat killed type IIIS bacteria by itself does not result in

any live bacteria culture, nor does type II mutate in to type III. A change or transformation of

type IIR into type IIIS must have occurred through the transfer of some active substance.

Grifffith’s results could be duplicated by mixing different types of heat killed and live-

bacterial strain in mouse (in vivo) as well by mixing item in test tubes (in vitro).

The search for the specific agent responsible for transformation, the transforming

principle continued until 1944. In this year Avery, Macleod and Mccarty showed that the

transforming principles consist entirely of DNA.

In their experiment they extracted the DNA from the heat-killed cells of type IIIS, and

mix this DNA extract directly with in vitro cultures of type IIR. A serum was added whose

antibodies react with the R cell and cause them to precipitate to the bottom. When transformation

occurs, type IIIS cells, not being precipitated, now grow diffusely throughout the medium.
Diagram showing Griffith’s experiment

Summary of Griffith’s experiment

Rough Bacteria in Mouse Mouse does not die

Smooth Bacteria injected in mouse Mouse die

Heat killed smooth bacteria injected in mouse Mouse does not die

Heat killed smooth bacteria +non heat killed Rough bacteria injected in mouse Mouse die.
Semiconservative DNA Replication

The particular mode of DNA replication suggested by Watson and Crick is called

semiconservative replication in which each partner strand of the duplex act as a template

directing the synthesis of a new complementary strand. The new duplexes are composed of one

parental and one new synthesized strand i.e the original duplex is partly conserved or semi

conserved in the new molecule.

This idea was compelling but it initially lacked experimental supports. In fact two other

possible modes of replication were suggested soon afterwards, the conservative replication and

dispersive replication. In the conservative replication, the original duplex remains intact i.e. it is

entirely conserved and the whole duplex guides the synthesis of a completely new duplex replica

of itself. In dispersive replication bits and pieces of newly synthesized DNA become assembled

with bits and pieces of the original duplex to reconstitute two duplexes from one original

templates.
 Diagram showing the three possible modes of duplex DNA replication. The predicted

distribution of original parental in purple color and newly synthesized strand in blue color

for two rounds of replication.

The nature of DNA

After the isolation and purification of the DNA molecule it was soon found that it molecular

weight varied widely. However it was established that DNA from all organisms are chemically

similar.

The DNA from any species consists of three chemical groups.

1. Phosphate

2 Deoxyribose sugar

3 Bases

There are two types of bases the pyrimidines (one ring compounds) and the purines (two ring

compounds). The pyrimidines are thymine (T) and Cytosine (C) while the purines are Adenine

(A) are guanine (G)

The DNA is a three dimensional structure that is accepted today and for which a nobel prize was

later awarded, was first proposed and demonstrated by J. D. Watson and Francis crick in 1953

(1953-1955). The DNA is a long molecule consisting of two strands. Each strand is a chain of

nucleotides. A nucleotide is a base plus a sugar. Successive nucleotides are linked together

through a phosphate group and a hydroxyl group on the sugar component. The two strands are

weekly associated by hydrogen bonds. The width of the two strands is always the same. This is

so because a purine (two ting structure) always pairs with a pyrimidine (one ring structure).

Specifically adenine always pair with thymine and Cytosine always pair with guanine.
 A+T
In any organism DNA molecule A=T and C=G. However the ratio varies widely for
G+ C

different species of DNA molecules

Since the above pairing is specific, if you know the nucleotide sequence of one strand you can

tell the sequence of the other strand. This phenomenon is referred to as complementarity.

Diagram showing Structure of the DNA

Ribonucleic Acid (RNA): RNA is also found as a component of the chromatin fibre

(Chromosome). Although both the DNA and RNA are nucleic acids. RNA differs from DNA in

a number of ways. The DNA is double stranded and has deoxyribose sugar.

The RNA is always single stranded with a ribose sugar. Both DNA and RNA contain the same

bases except that uracil is present in RNA in place of thymine which is found only in DNA.

RNA is synthesized in the nucleus on the DNA template transported into the cytoplasm where it

is more prevalent and associated with ribosomes. DNA on the other hand is self-duplicating and
exclusively located in the nucleus.

Several classes of RNA exist based on size and function. Heterogeneous nuclear RNA are very

large RNA molecules found in the nucleus. They undergo further processing before being

transported through the nuclear membrane into the cytoplasm as messenger RNA (mRNA) The

mRNA molecules carry the information from the gene in the nucleus to the ribosomes where

such information is translated into protein synthesis. Other classes of RNA include the smaller

transfer RNA (tRNA) and the ribosomal RNA (rRNA), which is found as integral parts of the

ribosomes. Both tRNA and rRNA play important roles in protein synthesis.

The genetic Code

One definition of the gene is that it specifies a peptide chain, through a complex procedure which

involves the transfer of information in a gene from the nucleus through a messenger (mRNA) to

the cytoplasm where the message is read during protein synthesis to make peptide chain. It is

possible to determine the base sequence code for an amino-acid sequence since there are four

bases in the mRNA. Since three base make a codon and a codon specifies an amino acid.

Crick and associates suggested a three letter word for the codon, so that there are 64 (4 3) possible

codons to the twenty amino acids. All these possible codons have ingeniously been assigned to

the amino acids for which they code.

Diagram for the base combinations (triplet codes) that specify different amino acids

Characteristics of the genetic codes

1. The genetic code is Universal (the same in all living organisms).

2. The code is degenerate since we have sixty four codes for only twenty amino-acids. This

means that one amino acids is coded for by more than one codon. The significance of this

is thought be a kind of stabilizer of species so that degenerate code reduces the effect of

base change mutations as the third of the codons are less important than the first two.

3 Codons are linear with no overlaps and only one base is part of only one codon.

4 The codon is directional and its is always read from a fixed point

5 In E. coli there is an initiation codon usually AUG at the beginning of all genes. There is

some evidence in support of AUG as initiation codon in higher organisms.

6 Termination codons in all systems are UAA, UAG and UGA.