Thursday, April 8, 2021

Gene Mapping, Two Point and Three Point Test Crosses, Coincidence and Interference

 Gene mapping

Genes are arranged linearly in a chromosome. The point in a chromosome where the gene is located is called locus. The diagrammatic representation of location and arrangement of genes and relative distance between linked genes of a chromosome is called linkage map or genetic map.

Constructing Genetic Maps

When two genes are close together on the same chromosome, they do not assort independently and are said to be linked.  Genes located on different chromosomes are not linked and they assort independently and have a recombination frequency of 50%.  Linked genes have a recombination frequency less than 50%.

The number of genetic recombinants produced is characteristic of the two linked genes involved. Genetic experiments can be used in genetic mapping.

Unlinked genes assort independently. The best cross to use to test for linkage is the testcross, a cross of an individual with another individual homozygous recessive for all genes involved. In a testcross between a+ /a b+ /b and a/a b/b, where genes a and b are unlinked, progeny phenotypic ratio will be 1:1:1:1. A significant deviation from this ratio (more parental types and few recombinant types) indicates that the two genes do not assort independently and that they  are linked. The chi-square test can be used to find the significant deviation.

In an individual that is doubly heterozygous for the w and m alleles, for example, the alleles can be arranged in two ways:


In the arrangement on the left, the two wild-type alleles are on one homolog and the two recessive mutant alleles are on the other homolog, an arrangement called coupling (or the cis configuration). Crossing-over between the two loci produces w+m and wm+ recombinants.

In the arrangement on the right, each homolog carries the wild-type allele of one gene and the mutant allele of the other gene, an arrangement called repulsion (or the trans configuration).  Crossing-over between the two genes produces w+m+ and wm recombinants.

The recombination frequency for two linked genes is the same, regardless of whether the alleles of the two genes involved are in coupling or in repulsion. Although the actual phenotypes of the recombinant classes are different for the two arrangements, the percentage of recombinants among the total progeny will be the same in each case.

In 1913, a student of Morgan’s, Alfred Sturtevant, determined that recombination frequencies could be used as a quantitative measure of the genetic distance between two genes on a genetic map. The genetic distance between genes is measured in map units (mu),where 1 map unit is defined as the interval in which 1 percent crossing-over takes place. The map unit is also called a centimorgan (cM), a term named by Sturtevant in honor of Morgan.

The unit of genetic map is Morgan or centimorgan. When the percentage of crossing over between two linked genes is 1 per cent, then the map distance between the linked genes is one morgan.

There is a greater probability of occurrence of crossing over, when the two genes are farther apart in a chromatid. The probability of crossing over between two genes is directly proportional to the distance between them.

When two genes are nearer, the probability of occurrence of crossing over between them is limited.

Gene Mapping with Two-Point Testcrosses

Testcrosses are used for mapping because the homozygous recessive parent produces only one type of gamete, with alleles that are recessive to the alleles in gametes produced by the heterozygous parent. So in a testcross we use one parent that is heterozygous for the genes being mapped and another parent that has the recessive alleles for those genes.

A two-point testcross should yield a pair of parental types that occur with about equal frequency and a pair of recombinant types that also occur with about equal frequency.

The following formula is used to calculate the recombination frequency:

Number of recombinants                 X 100    = Recombination Frequency  = Map Units

Total number of testcross progeny

The recombination frequency is used directly as an estimate of map units.

The two-point method of mapping is most accurate when the two genes examined are close together; when genes are far apart, there are inaccuracies.

Gene Mapping with Three-Point Testcrosses

Genetic maps can be built by using a series of two-point testcrosses. Still more complex type of mapping analysis for three linked genes can be done using a three-point testcross. In diploid organisms, the three-point testcross is a cross of a triple heterozygote with a triply homozygous recessive.

Uses of gene mapping 

1.        It is useful to determine the location, arrangement and linkage of genes in a chromosome.

 2.        It is useful to predict the results of dihybrid and trihybrid crosses.

Interference and Coincidence:

Besides single crossing over, having only one chiasma, there may be double or multiple crossing over. It has been discovered by H.J.Muller (1911) that when there are two double cross-overs (suppose a and b) then one cross-over (a) tries to prevent the formation of other cross over (b). This tendency of one cross-over to interfere with the other cross over is termed as interference. Suppose frequency of ‘a’ crossover is 10 and frequency of ‘b’ cross-over is 12, then their total frequency will not be 10+12 = 22 as required but will be less than 22 due to interference.

When the two things happen the same time and at the same place, they then coincide or intermix and this occurrence may be considered coincidence. This coincidence refers to the occurrence of two or more distinct cross-over (double or multiple) at about the same time in the same chromosomal region. Double cross-overs are the result of coming together (coincidence) of two single cross-overs.

When doubles occur in regular expected ratio, coincidence is said to be 100%, whereas interference will be nil. i.e., coincidence is inversely proportional to the interference.

According to Muller (1916) the coefficient of coincidence is the ratio between the observed and expected frequencies of double cross overs. 

Coefficient of coincidence = Actual number of double cross-overs/Expected number of double cross-overs.

Interference = 1- Coefficient of Coincidence

 

 

 

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