Chapter 15

Eukaryotic Chromosome Maping

 The discovery of Linkage
In the early 1900s Bateson
and Punnet were studying
inheritance in sweet pea.
They studied two traits
affecting the flower pod:
The color; P, purple and p,
red and the size of the
pollen grain; L, long and l,
round.
When PPLL was crossed
with ppll the F2 phenotypes
deviated strinkingly from
the expected 9:3:3:1
Bateson and Punnett Results
Phenotype Observed Expected
and 9:3:3:1
Genotype

P-L- 4831 3911

P-ll 390 1303
ppL- 393 1303
ppll 1338 435
 As an explanation, Punnett and Bateson
proposed that the F1 had produced more
PL and pl gametes than the recombinant
gametes Pl and pL.
Because these gametic types were the
parental types, the researchers thought
that physical coupling between the two
dominants P and L and the two recessive
p and l may have prevented the two genes
from independent assortment, however
they did not know the nature of the
coupling.
 Early Studies of Genetic Linkage: Morgans
Experiments with Drosophila

In Drosophila, the white eye gene w and a

gene for miniature wings m are located on
the X chromosome.
Morgan (1911) crossed a female white
miniature wwmm with a wild-type male
w+w+m+m+.
 In the F1, all males were white-eyed with miniature
wings (w m/Y), and all females were wild type for both
eye color and wing size w+wm+m.
F1 interbreeding in this case is the equivalent of a
testcross for these X-linked genes, since the male is
hemizygous recessive (wmY), passing on recessive
alleles to daughters and no X-linked alleles at all to
sons.
The use of the test cross is of extreme importance
here because one parent, the tester, carries only
recessive genes; therefore the phenotype of the
offspring reveals only the gametic contribution of the
other. Here one can concentrate on the meiosis of one
parent and forget about the other.
 In the testcross (F2 in this case), the most frequent
phenotypes for both sexes were the phenotypes of the
parents in the original cross (white eyes with miniature
wings, and red eyes with normal wings).
Non-parental phenotypes (white eyes with normal wings
or red eyes with miniature wings) occurred in about 37%
of the F2 flies. Well below the 50% predicted for
independent assortment, this indicates that non-parental
flies result from recombination of linked genes.
Genotypes
pr+- vg+- 1339
pr+ vgvg 1195
prpr vg+- 151
prpr/vgvg 154
2839
Obviously the genes deviate tremendously from the 1:1:1:1
expected and they indicate coupling of genes. The two
larger classes been the parental. The testcross also reveals
that the other two classes (non parental) are also found in
similar proportions.
Consider an opposite cross, where the parents are pr+pr+mm
and the second parent is ppm+m+. The F1 is pr+pr m+m.
Test cross the heterozygote (prprmm). The following
represent the results: Genotype
spr+- vg+- 157
pr+ vgvg 965
prpr vg+- 1067
prpr/vgvg 146
2839
Morgan proposed that:
The genes governing the same
phenotypes are located in the same pair of
homologous chromosomes. Thus when pr
and vg are introduced by the same parent,
these two genes are physically located in
the same chromosome. The same applies
for pr+ and vg+.
 Morgan suggested than the chromosomes pair during
meiosis and that they occasionally exchange parts in
a process called crossing over.

Final evidence for the physical presence of crossing

over was produced later on by Creighton and
McClintock who worked on corn and by Stern who
worked with D.m.
 Corn Experiments
Creighton and McClintock (1931)
presented the final physical evidence of
crossing over in Corn (Zea mays) plants in
which the two chromosomes under
study differed cytologically.
The study used a corn strain
heterozygous for two genes on
chromosome 9 . One gene determines
seed color (C for colored seeds, c for
colorless) and the other gene is involved
in starch synthesis. The wild-type allele
(Wx) produces amylose (the
combination of amylose and amylopectin
forms normal starch in corn seeds.) The
waxy mutant (wx) lacks amylose, and has
waxy starch containing only amylopectin.
These two genes C and Wx are linked.
 Summary of Genetic Maping
Genes on non-homologous chromosomes
assort independently, but genes on the
same chromosome (syntenic genes) may
instead be inherited together (linked), and
belong to a linkage group.

In order to determine linkage, one analyzes

the frequency of allele recombination in
progeny of genetic crosses.
 a. If the genes are linked the gametes stay as their
parents. These are klnown as the parental.
b. Gametes with new associations of parental alleles
produced by crossing
over or genetic
recombination are
called the
recombinants.
 c. Testcrosses determine which genes
are linked, and a linkage map (genetic
map) is constructed for each chromosome.
 d. Genetic maps are useful in
recombinant DNA research and
experiments dealing with genes and their
flanking sequences.

e. Current high-resolution maps include

both gene markers from testcrosses, and
DNA markers composed of genomic
regions that differ detectably between
individuals.
 Linkage symbolism
prvg This is the same than pr+pr vg+vg and it is
pr+vg+ said to be in coupling or cis position. The
linkage group is prvg and pr+vg+

pr+vg This is the same than pr+pr vg+vg and it is

said to be in repulsion or trans position.
prvg+ The linkage group is pr+vg and prvg+
 Constructing a Genetic Map
A linkage map shows the physical location
of the linked genes in a chromosome.
A linkage map uses the percentage of
genetic recombination as a quantitative
index for the linear distance between the
two genes on a genetic map.
A recombination frequency of 0.01 = 1
map unit or 1 cM (centimorgan).