Chapter 8

Genetic Recombination
Have you ever played the lottery?
 Why are we so diverse?

1. Mutation

2. Random Fertilization

3. Recombination

Why is it important to
be diverse?
Mechanism of Genetic
Recombination
a. Requires 2 DNA molecules that similar but non-
identical

b. Homology allows DNA on different molecules to line

up and recombine precisely

c. Enzymatic cutting and pasting of both DNA

backbones from each of 2 DNA molecules required
for recombination

So what kinds of DNA are similar but not identical?

REMEMBER…

Homologous chromosomes
Simplified Model of Genetic
Recombination
Homologous
2 similar but
nonidentical DNA

Enzymatic
‘Cut and Paste’

2 Recombined DNA
Genetic Recombination in Bacteria

a. Genetic recombination occurs in E. coli

b. Bacterial conjugation brings DNA of two cells

into close proximity

c. Transformation and transduction provide

additional sources of DNA for recombination
Genetic Recombination in Bacteria

• Some bacteria genetically reshuffle

– Genes transferred from one individual to
another recombine with existing DNA

How did they figure this out?

– Genetically identical clones allow molecular
genetics studies
 Genetic Recombination in E. coli
• Prototrophs - Bacteria grow on minimal media because they make
their own amino acids

• Auxotrophs – Bacteria with mutations does ____ grow on minimal

medium

– Three letter gene name:

– +
normal, – mutated allele
– Ex. arg+ and arg¯

Complete medium - full complement of nutrients

Minimal media – missing some nutrients

 Replica Plating
• Replica plating – is a technique that is used to:

1. identify prototrophs versus auxotrophs

2. identify and count genetic recombinations in bacterial

colonies
 Replica Plating

Which are
prototrophs
and which are
auxotrophs?

Prototrophs
Prototrophs
only
and
auxotrophs NO
Auxotrophs
 What is the experimental evidence for
Genetic Recombination in Bacteria?
Lederberg
& Tatum

But how
does this
happen??
1. Bacterial Conjugation
• Bacterial recombination by conjugation
– Bacteria are haploid
– ________ connects two bacteria
– Donor sends DNA via cytoplasmic bridge to recipient

• Recipient undergoes recombination

– Plasmids: Circular, nonchromosomal transferable
DNA
– R plasmids confer resistance to antibiotics

__________ Gene Transfer

Conjugating E. coli Cells
F Factor and Conjugation
• Donor cell must have ________(fertility)
plasmid
– F+ cells are donors with F factor
– F- cells are recipients without F factor

• F factor has genes that encode for sex pilus

– cytoplasmically connects F+ cell to F- cell
– F- cell converts to F+ cell
– ___ recombination occurs
Transfer of Genetic Material
During Conjugation
Bacterial
chromosome

Rolling Circle Replication

Yes F plasmid is transferred

___ chromosomal DNA transferred

___ DNA recombination

 Hfr Cells and Recombination
• Hfr (high frequency) cells integrate F factor into
bacterial chromosome through recombination
– Hfr cells can conjugate with F- cells
– Recipient becomes _________

• Genetic recombination occurs by double crossing-

over in recipient
– New generations have __________
 Transfer of Genetic Material
During Conjugation

Partial Diploid

___ partial
chromosomal DNA
F factor into transferred
chromosomal
DNA = Hfr cell ___ DNA
recombination
Mapping Genes by Conjugation
• Mated Hfr and F- cells that differed in number
of alleles

• At regular intervals after conjugation

commenced, remove cells and break apart
mating pairs

• Cultured separated cells and analyzed for

recombinants
Mapping Genes by Conjugation
• Greater time to conjugate before separation,
the greater number of donor genes into
recipient
 greater number of ________

• Note order and time at which genes were

transferred
--> able to map and assign relative positions of
several genes of E. coli chromosome
 2. Transformation
• Transformation occurs when bacteria take up DNA
from disintegrated bacteria

– Linear fragments recombine by double crossovers

– Transformation bacteria usually have DNA binding
protein in wall

• Artificial transformation
– Alters cell membrane for DNA penetration
– Electroporation
_______Gene Transfer
 3. Transduction

• Transduction occurs when bacterial phages

transfer DNA from one bacteria to another

• Virus incorporates DNA fragments from host cell

– If DNA fragments are homologous
 bacteria become partial diploid

– Recombination by double crossovers

________Gene Transfer
 3a. Generalized Transduction

Phage Proteins
Phage Attachment

Phage Assembly
Phage Enzymes

Phage Release

Phage DNA Replication

Phage with
_______DNA
Recombination can now occur
3b. Specialized Transduction

Prophage

Fig. 10-8, p. 208

Virulent vs. Temperate Bacteriophage
• Virulent Bacteriophage – uses only lytic cycle
of infection
 ____ host bacteria

• Temperate Bacteriophage – uses both lysogenic

and lytic cycle of infection
 may or may not kill host bacteria

• Prophage – bacteriophage integrated into host

DNA
 Genetic Recombination in
Eukaryotes: Meiosis
a. Meiosis occurs in different places in different
organismal life cycles

b. Meiosis changes both chromosome number and

DNA sequence

c. Meiosis produces four genetically different

daughter cells

d. Several mechanisms contribute genetic diversity

 Sexual Reproduction

• Sexual reproduction produces offspring by union of male

and female gametes (sperm and egg)

– Meiosis produces gametes with half chromosome

number

– Evolutionary advantage: __________

 Fertilization

• Fertilization fuses nuclei of egg and sperm

 Zygote

– Restores parental chromosome number

 Animal Life Cycles
• Diploid phase dominates animal life cycles
– Meiosis followed directly by gamete formation
– Haploid phase is reduced and short, no mitosis

• In males, all four nuclei from meiosis form

separate sperm cells

• In females, only ______ nucleus becomes an

egg
 Homologous Chromosome Pairs
• Paternal chromosomes from male parent
• Maternal chromosomes from female parent

– Homologous chromosome pairs

– Alleles may be different within homologous pairs

• Meiosis separates homologous pairs

– Before meiosis - diploid (2n)
– After meiosis - haploid (n)
 Meiosis I

• Meiosis I: First meiotic division

– Recombination exchanges segments between
homologues
– Produces two haploid cells with chromatids
attached
 Meiosis II

• Meiosis II: Second meiotic division

– Sister chromatids separate into separate cells
– Produces 4 recombined haploid cells
2 Meiotic Divisions Give 4
Haploid Nuclei
Diploid

Sister Chromatids

Recombination

Haploid Non-identical
 Meiotic Cell Cycle
• Prophase I - Sister chromatids condense Recombination
chromosomes

– Synapsis Synapsis
pairing of
homologs

– Tetrads
 fully
paired homologs

– Recombination 
mixes alleles
across tetrads
Meiotic Cell Cycle

• Prometaphase I
– Nuclear envelope
breaks down
– Kinetochores attach to
polar spindles
Meiotic Cell Cycle
• Metaphase I and
Anaphase I
– Tetrads align on
metaphase plate
– Homologs segregate -
move to poles (sister
chromatids attached)

– Nondisjunction creates
abnormal chromosome
numbers Random
Alignment
 Meiotic Divisions
• Telophase I and Interkinesis
– NO change in chromosomes
– Spindle disassembles

Fig. 10-11, p. 214

Meiotic Cell Cycle (cont’d)
• Prophase II,
Prometaphase II
– Chromosomes
condense, spindles
form
– Nuclear envelope
breaks, kinetochores
attach to
microtubules
Meiotic Cell Cycle (cont’d)
• Metaphase II
– Chromosomes align
on metaphase plate

Random
Alignment
Meiotic Cell Cycle (cont’d)
• Anaphase II and
Telophase II
– Spindles separate
chromatids
– Spindles disassemble
– New nuclear
envelopes form

___ genetically different

haploid cells
 Nondisjunction
• Both members of pair of homologous chromosomes
connect to spindles from same pole

• Following anaphase, one pole then receives both copies

of pair, and other pole receives none

• Result is gametes that have 2 copies of a chromosome

• After fertilization, zygote has 3 copies of chromosome

instead of 2

• Ex. Trisomy 21
 Sex Chromosomes in Meiosis

• Meiosis and sex chromosome inheritance:

– Gametes produced by females may receive either X

chromosome

– Gametes produce by males may receive either X or Y

chromosome
 Meiosis and Mitosis Compared

• Mitosis and meiosis compared

– Both: Similar cell divisions, meiosis divides twice
– Mitosis: Two identical daughter cells (diploid)
– Meiosis: Four genetically different cells (haploid)

• Premeiotic interphases similar to mitotic

interphase
– Chromosomes copied into sister chromatids
 Comparison of Key Steps in
Meiosis and Mitosis

Random
Alignment

2 identical
diploid cells
4 non-identical
haploid cells
Genetic Variability
Four main ways:

1. Genetic recombination

2. Random segregated at anaphase I

3. Alternative combo at anaphase II

4. Random fertilization
 1. Genetic Recombination

• Recombination (_________)
– Key genetic shuffle of prophase I

• Tetrads held together at synaptonemal complex

– 2 of 4 chromatids exchange alleles
– Chiasmata or crossovers are points of exchange
Crossing-Over
Where does this happen?

Between non-sister chromatids

Synaptonemal Complex
 2. Random Segregation

• Random segregation
– Key genetic shuffle of metaphase I

• Each chromosome of a homologous pair may

randomly end up at either spindle pole
– Any combo of maternal and paternal
chromosomes  segregated to gametes
– 2X number of possible combinations
 Random Spindle Connections

At Metaphase I

Chromosomes
line up
randomly
3. Alternative combo at anaphase II

At Metaphase II

Attachment of spindle
to kinetochore on sister
chromatids is random

Therefore alignment is
random.
 4. Random Fertilization
• Random chance of male and female gamete
forming zygote

• Meiosis allows randomness necessary for

Mendelian laws of inheritance
Putting it into perspective

1.How is diversity of genetic material

advantageous?

2.What are the mechanisms in prokarytes to

achieve genetic diversity?

3.What are the mechanisms in eukarytoes to

achieve genetic diversity?
http://www.youtube.com/watch?v=D1_-
mQS_FZ0