Chapter 4 question 7 the answer key is WRONG

The answers should be:

Initial cross AB X ab
or A/A . B/B X a/a . b/b
meaning A and B are the two genes linked
F1: AaBb

F1 testcrossed: AaBb X aabb

a. 25 %

b. 50 %

c. 45%

d. 38%
 Summary from last lecture…

Finished mapping 3 linked genes in relation to one another

The number of double cross over events can be inaccurate if there was
interference
Complete interference (I=1) we see no double crossovers

No interference (I=0) we see all the double crossovers we expected

Mapping a centromere
– use Neurospora since it gives us ordered meiosis products

Mapping genes using tetrad analysis – figure out parental, non-parental and
tetratype classes of tetrads
 In Drosophilia
Crossing over ONLY occurs in females So any crossing over we see came from one
parent only and makes mapping easier
Testcross a female heterozygous for Ly Sb and br to a tester male
You know that Ly and Sb are located on chromosome 3
Data: Phenotype #
Ly + br 404
Parental no crossover
+ Sb + 422
Ly Sb + 18
Single cross over – Ly is different
+ + br 16
Ly Sb br 75 Single cross over – Sb is different
+ + + 59
Ly + + 4
+ Sb br 2 DOUBLE cross over
– br is different
Total observed 1,000
So, the order is: Ly---br---Sb The parental has
Ly, br and +(sb+) linked
Ly to br distance is: (18 + 16 + 4 + 2 = 40) or 4 m.u.s &
Ly+, br+ and sb linked
Sb to br distance is: (75 + 59 + 4 + 2 = 140) or 14 m.u.s
 Another practice problem

In Drosophila a cross was performed involving the abc phenotypes:

We get the following phenotypic ratios for the offspring:

Abc 460 No cross over 91% What’s the order of these genes?
aBC 450 We think b is in the middle
abc 32 But no double crossovers
Single cross over (a is different) 7%
ABC 38 Exist to confirm this
aBc 11
single cross over (c is different) 2%
AbC 9

Total observed: 1,000

What phenotype is not seen? ABc or abC where b is different
NO DOUBLE CROSSOVER OCCURRED
A and B are 7 m.u.s apart
B and C are 2 m.u.s apart

How many double crossovers did we expect to see? (0.02 X 0.07 X1000) = 1.4

What is I here? I= 1- c.o.c. = 1-0/1.4 I= 1-0 or 1 Complete interference

 End of chapter 4

Omit: section 4.4, 4.5, 4.6 (except the Perkins formula)

pages 132 -135
 Bacterial genetics
Bacterial chromosome

Many bacteria only have 1 circular chromosome

This chromosome is unlike eukaryotic chromosomes

it does not wrap around histones and form nucleosomes
it supercoils to become compact so it fits into a cell

http://web.siumed.edu/~bbartholomew/images/chapter29/F29-17.jpg
E.Coli is a
widely used
organism in
genetics
-Small & easy
To keep in a lab

-Quick
reproduction
time

-Asexual
reproduction
Or conjugation

-easy to
manipulate

-easy to
introduce &
keep foreign
DNA in the cells
 Only one copy of
each gene in the
Fission produces cell’s genome
New individuals
Genetically
identical
To the original
parent cell
 Bacterial growth
Normal bacteria can grow on minimal media & these are called prototrophic organisms
minimal media – give cells glucose and some inorganic salts to live on
Mutant bacteria may not be able to grow on minimal media – these are auxotrophic
Grow on rich media certain essential biomolecules must be present
– the cell can’t make them all

Bacteria can
Grow in solution
Or on a solid
Agar gel
Surface

Visible colonies
Contain identical
cells

The ability of a bacterial cell to grow in minimal media versus “rich” media is a
phenotype that we can observe
This allows us to tell the difference between bacterial strains & this is referred to as the
cells’ having genetic markers
 An example of what E. coli looks like growing on
Agar media (a spongy surface cells can grow on)

An example of a plasmid

Colonies are an area of visible cell growth containing Colonies we see as

cells which are genetically identical to one another green cells have a
gene that
Source: http://www.bio-rad.com/B2B/BioRad/product/ makes green
fluorescent protein
 Bacteria can share genetic information
There are three main ways that bacteria can obtain new DNA (or some genetic
variation)

- Conjugation – a parasexual process where genetic information is passed from one

bacteria to another bacteria (only a one-way transfer)
Here two bacterial cells must physically contact one another

- Transformation – small pieces of DNA in the environment are taken into a bacteria

-Transduction – transfer of genetic material between bacterial cells that

Is mediated by bacteriophages
Bacteriophages or phages – are viruses that attack bacteria
DNA from an infected cell is taken into a new viral particle that goes and
infects a new cell and thereby gives the new cell some genetic
information from the cell that was infected first
For all 3 of these processes the end result is the same: a bacterial cell takes in
and makes some foreign DNA part of its genome
 Conjugation
DNA is transferred from one cell to another THIS IS A ONE-WAY TRANSFER OF DNA
DNA moves from an F+ cell into an F- cell
Donor cells have a fertility factor – they are F+
These donor cells make the pili (or pilus)
Recipient cells do not have a fertility factor and are F-

This fertility factor gene can be located:

- on the bacterial chromosome (that makes these bacteria Hfr cells) or
- on another piece of DNA in the cell that is known as a plasmid (shown in red)

Can be an F+, Hfr or F’ strain

Is initially an F- strain
 Hfr strains
How the strain is made –
a double crossover event between the plasmid DNA and bacterial chromosome
the end result is that the plasmid becomes incorporated into the chromosome
at some insertion point
 Making an F’ strain
Start with an Hfr strain and the F factor region recombines in order to
leave the bacterial genome and become a separate plasmid
When this recombines out of the genome often some of the bacterial genes will end
up on the new F factor plasmid (here the lac gene is shown being removed
from the bacterial chromosome and put into the F factor plasmid

Note: this is
A reversal of
What
happened
When we
made the Hfr
strain

Again remember an F+ strain can be called:

F+, Hfr or F’
 Diagram of bacterial conjugation

A cut in the middle of the F factor initiates the single-strand of

DNA that is transferred

Donor 2 crossing Linear

Replicates Over events Chromosomes
Its genome But the whole allows for are lost when
F factor is not donor and the bacterial
present initially recipient cell divides
In the recipient DNA to be
exchanged
The complete F factor is always the very last portion of the donor chromosome to
be transferred
Once complete parts of the donor genome enter the recipient cell the DNA can
integrate and become part of the recipient’s genome making the recipient an
exconjugate.
 Diagram of bacterial conjugation (con’t)

Here the recipient cell

Is a partial diploid

Also called a
- merozygote

NOTE: after conjugation the donor’s genome is UNCHANGED

only the recipient’s genome can change
 What type of information can a conjugation experiment tell me?

Let an Hfr (or F+)and F- strain conjugate for different amounts of time & this can
tell us what genes are close together and in what relative order genes are
on the donor bacterial chromosome

A F
For red arrow A B C D F is the order of transmission
D
B C For black arrow D C B A F is the order of transmission
Donor chromosome
Each triangle is an origin (O) –
The origin is where: the nick is made, the single-stranded DNA for transfer starts
& the copying donor genome starts
The origin is the first bit of donor DNA that shows up in the recipient cell
Different donor strains will have the F factor in a different location and a different
orientation (as shown above)
When studies compared different F+ strains the results where not 100% identical
It terms of the actual order of genes that entered the cell
BUT
When compared the different strains appeared to have genes transferred into the
donor in similar patterns
Different bacteria will have the F-factor in a different position of orientation with
relation to one another
Diagram of two different bacterial genomes

Notice: the actual

order of the donated
genes differs BUT
the pattern does not

Here the order of genes is leu first

Here, leu is next to thr
Here, thr is next to thi

Here the order of genes is thi first

Here, leu is next to thr
Here, thr is next to thi
 How can I develop a chromosomal map from bacterial conjugation?

We use an interrupted mating experiment

here specific bacterial strains are allowed to conjugate for various lengths
of time.

With a very short mating time only 1 or 2 genes may transfer – these genes
will be close to the origin

With a long mating time more genes will transfer

F factor will always transfer last

To get a map distance between genes we use the time between the transfer of
any 2 genes as the distance between those 2 genes
so, Map units = minutes

To determine map units between two genes compare the times

when genes FIRST appear
 Using conjugation how do I map a bacterial chromosome?

A sample problem:
an interrupted mating experiment was done using the following strains
Xyl+, leu+, mal+, his+ and F+ to a Xyl-, leu-, mal-, his- and F-
The table below is displays the phenotype information for the recipient strain
Time
Determine map units by comparing the times
Of mating
when genes FIRST appear
Minutes % phenotype
5 25 xyl+ Now:

10 80% xyl+ O and xyl are ~ 5 m.u.s apart….

15% mal+ 1 minute = 1 map unit

15 90% xyl+ 5 m.u.s 5 m.u.s 15 m.u.s

75% mal+

20 90% xyl+ O xyl mal his

80% mal+
Note: we are looking for a change in the
25 90% xyl+
phenotype of the F- strain in comparison
80% mal+
to the phenotype of the strain at
20% his+
time = 0 minutes (before mating begins)
 The following data is the analysis of the recipient bacterial strain during an
Interrupted mating experiment between a leu+ pro+ arg- F+ and a leu- pro- arg+
F- strain.
What is the order of the genes on the donor chromosome and the appropriate
map distances?
%
Time Of
(minutes) Phenotype of recipient strain
cells
DNA moves one way
2.5 100 arg+

5 100 arg+
9 pro+ F+ F-
8 86 arg+ Leu+ Leu- At time
50 pro+ Pro+ Pro- 0 min
Arg- Arg+ The F-
15 15 arg+ Cells
88 pro+ Are arg+
Donor recipient
So, we are looking for a decrease in arg+ cells
as our difference at the arg gene
Order: Origin pro arg leu F
O & pro are 5 map units apart, O and arg are 8 map units apart,
arg and pro are 3 map untits apart
 Transformation
A process through which bacteria can take DNA up from its environment
this DNA maybe different than anything that is in the recipients genome
and this foreign piece of DNA may integrate, that is recombine with
the recipient’s genome and become part of the recipient’s genome.

This DNA in the environment is usually left over from another cell that as died
Transformation occurs with pieces of DNA that are small (10,000 to 20,000 bases long)
E. coli chromosome has about 2,000,000 bases
 End of chapter 5

Omit the following sections:

Fine-scale mapping page 161-162
R plasmids page 164 – 165
Mapping phage chromosomes pages 168 - 170
specialized transduction part pages 172 – 174
Physical maps versus linkage maps pages 174 – 175