Three-Point Test Cross

A three-point test cross is a technique used in genetic mapping when considering the inheritance of
three alleles. It can be used to order three loci on a chromosome, and map the distance between these
loci in centimorgans (cM). The centimorgan is equivalent to the frequency of recombination; a map
distance of 1cM across a chromosome is equivalent to a recombination frequency of 1% for that
chromosomal region.

The three-point test cross involves a mating between a triple heterozygote and a triply recessive
homozygote. Once offspring phenotypes have been identified and counted in classes, the procedure is
as follows:

Sum the offspring to get a total number of progeny

Identify the offspring with parental genotypes (these will have highest frequency classes)

Identify the offspring with double recombinant genotypes (these will have the lowest frequency classes)

Identify which locus has swapped position, relative to the other two, in the double recombinant
genotypes compared to the parental genotypes - this locus is the middle locus on the chromosome, and
the other two loci may be placed either side (in any order)

Draw a map of the chromosome, and divide the spaces between loci into two 'regions'

For the first region identify the two classes of single recombinant offspring where a recombination event
has occured in this area. Sum the offspring of these classes and the offspring of the double recombinant
classes to give you a recombinant value for that region

The map distance for this region is then (recombinant value)/(total number of progeny) x 100

Repeat steps 6 & 7 for the second 'region' to give this region a map distance

Three Point Test Cross: Multiple Point Gene Mapping

Gene mappers are motivated to map all of the tens of thousands of genes found on the chromosomes of
plant or animals. Analyzing data from crosses to determine map distances for two genes at a time makes
the process time consuming and tedious. Therefore geneticists will often attempt to map as many genes
as possible from a single set of progeny. We will go through the simplest multiple point mapping
example, a three point testcross, to demonstrate the process.

We will use our three corn seed trait loci again and use data from one cross to map these three loci. The
first step would be to obtain a trihybrid individual that is heterozygous at all three loci and then perform
a testcross with this trihybrid.

In genetics, the coefficient of coincidence (c.o.c.) is a measure of interference in the formation of

chromosomal crossovers during meiosis. It is generally the case that, if there is a crossover at one spot
on a chromosome, this decreases the likelihood of a crossover in a nearby spot.[1] This is called
interference.

The coefficient of coincidence is typically calculated from recombination rates between three genes. If
there are three genes in the order A B C, then we can determine how closely linked they are by
frequency of recombination. Knowing the recombination rate between A and B and the recombination
rate between B and C, we would naively expect the double recombination rate to be the product of
these two rates.

The coefficient of coincidence is calculated by dividing the actual frequency of double recombinants by
this expected frequency:[1]

c.o.c. = actual double recombinant frequency / expected double recombinant frequency

Interference is then defined as follows:[1]

interference = 1 − c.o.c.

This figure tells us how strongly a crossover in one of the DNA regions (AB or BC) interferes with the
Interference And Coincidence

Just before crossing over actually occurs, an event called synapsis (pairing of homologous chromosomes)
occurs during which chiasma forms. Chiasma is a (X-shaped) structure formed when non-sister
chromatids from homologous chromosomes overlap, as a prelude to recombination when DNA is
exchanged. Because one chiasma can cause stearic hindrance to the formation of another chiasma
within its close proximity, double crossovers do not occur at the expected frequency. This phenomenon
is called interference.

Interference refers to the reduction in the probability of occurrence of crossover at one location on a
chromosome due to crossover occurring in another location.

Let us assume that there are three genes, x-y-z, lined up in a chromosome. If the recombination value
for x-y is 20% and for y-z is 40%, and if they are independent of each other, then the recombination
value of the double crossover value between x and z should be 20% of 40%, which should be 8%.
However, in reality, this has always been lower, and it happens because of interference.

The degree of interference varies from location to location. If double crossover does not occur at all, it
would mean that interference is 100%. If double crossover occurs at the expected frequency, then it
would mean that interference has not occurred at all.

Coincidence is another term to express the same phenomenon, and is the complement of interference.
If double crossover occurs at the expected frequency, then coincidence would be 100%, and if double
crossover does not occur at all, then coincidence would be 0%.

In the example illustrated above, the expected double crossover is 0.2 × 0.4 = 0.08. If the observed value
of double crossover is 2%, then the coefficient of coincidence would be 25%. The greater the
coincidence, the lesser is the interference.

Although positive interference should be expected to be common, negative interference occurs in some
bacteria and fungi. In these organisms, recombination in one location enhances the occurrence of
recombination in another location.formation of a crossover in the other region.

Little is known about the factors that influence the frequency and distribution of meiotic recombination
events within human crossover hotspots. We now describe the detailed analysis of sperm
recombination in the NID1 hotspot. Like the neighbouring MS32 hotspot, the NID1 hotspot is associated
with a minisatellite, suggesting that hotspots predispose DNA to tandem repetition. Unlike MS32,
crossover resolution breakpoints in NID1 avoid the minisatellite, producing a cold spot within the
hotspot. This avoidance may be related to the palindromic nature of the minisatellite interfering with
the generation and/or processing of recombination intermediates. The NID1 hotspot also contains a
single nucleotide polymorphism (SNP) close to the centre, which appears to directly influence the
frequency of crossover initiation. Quantitative gene conversion assays show that this SNP affects the
frequency of gene conversion and crossover to a very similar extent, providing evidence that
conversions and crossovers are triggered by the same recombination initiating events. The
recombination-suppressing allele is over-transmitted to recombinant progeny, and provides the most
dramatic example to date of recombination-mediated meiotic drive, of a magnitude sufficient to
virtually guarantee that the recombination suppressor will eventually replace the more active allele in
human populations.