II.

Sex and the Single Cell

A. Prokaryotes
1. Characteristics
a. Have no nuclear membrane.

b. Have few membranous organelles.

c. Have a single circular DNA or “chromosome” and sometimes

plasmid DNA molecules.

d. DNA molecules are “naked” having little protein associated with

the molecules.
2. Theta () Replication

a. Replication is asexual and daughter cells are identical to the

parental cell.

b. Replication initiates at a single replication origin and proceeds

bidirectionally (two forks).

c. Both chromosomal and plasmid DNA’s carry out  replication

under normal circumstances.

d. Plasmid DNA’s must have a replication origin or the bacterial

cell’s replication machinery will not replicate the plasmid.
B. Gene transfer in prokaryotes
1. Detection of gene transfer in prokaryotes depends
on the use of metabolic mutants.
a. Auxotrophs - require a metabolite not needed by the wild type
prototroph. Example: Arg+ vs Arg -

b. Antibiotic resistance - have ability to grow in the presence of an

antibiotic. Example: AmpS vs AmpR

c. Utilization mutants - different from wild type by having

the ability or inability to use certain carbon or nitrogen sources.
Example: Lac+ vs Lac-
Replica Plating
 2. Transformation - DNA mediated gene transfer

a. Competent recipient cells bind and take up DNA.

b. DNA is integrated into recipient genome and the displaced strand is

degraded. This action is mediated by the Rec A protein and results in a
homologous recombination event.

c. Replication of heteroduplex DNA can form homoduplex transformant.

d. Some bacteria are naturally competent; S. pneumoniae.

e. Bacteria can also be made competent to take up DNA fragments or

plasmids.
R R’ S’ Single-strand invasion and displacement

S’

R
R’ R S’

Replication
R R’
R S’ S’ S’

Transformant
3. Conjugation I – Infective Conversion

a. Some E. coli have a specialized plasmid called the

F factor (F+). The F factor encodes proteins
required for transfer of the F factor to cells lacking
the plasmid (F-).

b. Among these proteins encoded by the F factor are

those that form the pili. These hair-like structures
establish contact with the F- cell and upon
adherence, lead to the formation of the conjugation
bridge.
Figure 7.3 top
c. The transfer of the F factor is carried out by rolling circle
replication (also called  replication).
 Figure 7.3 bottom

d. Cells receiving the F factor are now F+. This process

is called infective conversion. No genomic DNA is transferred.
 4. Conjugation II - Cell mediated gene transfer.

Figure 7.11

a. Hfr (high frequency of recombination) cells are formed when

the F factor integrates into the bacterial genome.
 Conjugation II - Cell mediated gene transfer
continued.

b. When these cells attempt infective conversion bacterial

genes are frequently transferred along with part of the F
factor.

c. The conjugating pair usually separates before the entire F

factor is transferred so the recipient cell rarely becomes F+.

d. However, the transferred bacterial genes can recombine

with the recipient cell’s DNA and cause a change in
genotype.
Hfr leu+tetS tetS gene F- leu-tetR

leu+ gene

Fig. 7.12 Top. Hfr x F- conjugation

 Hfr leu+tetS

To identify the leu recombinant

+ leu+ leu-
the exconjugants must be grown
on minimal media containing
tetracycline (counter selection
against the original Hfr cell).
leu+

F- leu+tetR
Fig. 7.12 Bottom. Hfr x F- conjugation
Figure 7.14

Purple arrows indicate the

map position of known Hfrs
5. Transduction - Phage mediated gene transfer

a. Transduction is carried out by temperate bacteriophage. These

phage can follow two pathways, the lytic or the lysogenic
pathway.

b. In the lytic pathway, the phage injects its DNA into the
bacterium that now replicates the phage DNA. The phage DNA
assembles with the phage coat proteins and then lyses the cell.

c. The lysogenic pathway occurs when the phage DNA becomes

inactive or latent. The DNA either remains a circular plasmid or
integrates into the bacterial chromosome. This inactive form is
called the prophage and the cell plus the prophage is called a
lysogen.
Lytic Pathway
Lysogenic Pathway
 Transduction Continued

d. Prophage induction can occur when lysogenic cells are

stressed. At this time the bacteriophage DNA becomes active
and begins replication. New phage particles are produced and
lysis occurs.

e. During packaging bacterial DNA may be inadvertently

inserted into the phage head. These particles are called
transducing phage.

f. Transducing phage or particles can inject the bacterial DNA

into another bacteria. If the injected DNA is integrated into
the recipient bacteria’s chromosome the genotype of the
bacteria may change.
III. Eukaryotic Cell Division
A. Eukaryotic chromosomes - Linear, gene bearing
units of inheritance.
1. Chromatin is composed of DNA and proteins.
a. Small, basic proteins called histones bind tightly to the DNA as
an octamer (8 individual proteins in a complex).

b. Binding of histones to DNA causes coiling of DNA around

octamer. This forms a structure known as the nucleosome and
causes compacting of the DNA. Additional, non-histone proteins
help to further compact the DNA.
Compacting of DNA
 2. Structural features of chromosome.

a. Double-stranded DNA having multiple origins of replication.

b. Centromere - constricted region of chromosome to which spindle

fibers attach during mitosis and meiosis. Also point at which
replicated chromosomes (sister chromatids) are connected prior to
separation.

c. Telomere - special repeated sequence at the ends of the chromosome.

 B. Cell Cycle

1. G1 - growth of the cell

2. S Phase - replication of DNA
3. G2 - Final preparations for cell division
4. M Phase - mitosis, actual separation of the
replicated chromosomes and division of
cellular components into two cells.
 C. Mitosis
1. Prophase

a. Condensation of chromosomes.
b. Nuclear membrane disappears.

2. Metaphase

a. Sister chromatids line up at axis of cell.

b. Spindle fiber apparatus forms and attaches to the centromeres of the

sister chromatids.

c. Centrioles appear at the poles of the cell and are also attached to
spindle fibers.
 Mitosis Continued
3. Anaphase

a. Contraction of spindle fibers pulls sister chromatids apart. There is

now one copy of each original chromosome migrating towards the
poles of the cell.

4. Telophase

a. Nuclear membrane reforms around the chromosomes and two

daughter cells are formed as cell membrane forms between the two
compartments.
IV. Eukaryotic Life Cycles
A. Asexual Reproduction
1. DNA simply replicates generation after
generation (mitosis).
B. Sexual Reproduction
1. Haploid - Having only one copy of each chromosome.
a. Haploid gametes fuse to form a 2N zygote which undergoes
meiosis (similar to mitosis but with two divisions of the nuclear
contents) to form haploid meiotic products. These products will
now grow and divide by mitosis until they in turn act as gametes.
 Gist Fig 3-1
Haploid parents A a

Mitosis

Haploid gametes A a

Fertilization

Diploid Zygote Aa

Meiosis and segregation

Haploid meiotic A A a a
products
2. Diploid - Having two copies of every chromosome.

a. Meiosis takes place in the specialized germ cells (oocytes and

spermatocytes) of adult individuals. The 1N products of this division
become eggs and sperm in animals and ovules and pollen in plants.

b. These specialized gametes fuse to form a diploid zygote which

matures into the diploid adult.
 Gist Fig 3-1
Diploid parents AA aa
Meiosis
Haploid A A AA a a a a
gametes
Fertilization
Diploid zygote (F1) Aa

Mitosis

Diploid Adult (F1) Aa

Meiosis

Haploid gametes of A A a a
the F1
3. Haplo-diploid

a. Both haploid and diploid vegetative stages can multiply by mitosis.

b. Example - Yeast Life Cycle

1N a Mitosis  Asexual reproduction (budding)

 
2N a/
 
1N  Meiosis  4 1N products of which 2 are a and 2 are 