Objectives Unit Six:  Linkage and Chromosome Mapping

1.  Define the following terms and utilize these terms in an appropriate context:
Chiasma                      chromosome theory                  complete linkage            crossing over
double crossover        linkage                                        linkage group
coupling                       repulsion                                     cis/trans                           parental group
crossover group

2.  Demonstrate ways to indicate a genotype involving linked genes.

3.  Using chromosome diagrams and gene symbols (genotypes), show how the offspring
    resulting from a test cross of individuals heterozygous for two traits could be mainly of two
    phenotypic classes (demonstrating linkage but no crossovers).

4.  Explain how it could be shown that the results of the cross referred to in objective #3 were
    not caused by any affinity of dominant gennes for each other, or recessive genes for each other.

5.  Explain a crossover event and illustrate the resulting offspring of such an event.

6.  Explain why more crossing over occurs between two distantly linked genes than between
    two genes that are very close together on the same chromosome.

7.  Explain why double-crossover events are expected in lower frequencies than single-crossover
    events.

8.  In general terms, give the relationship between the spacing of genes on a chromosome and
     the crossover frequency;  and state specifically the relationship between crossover frequency
     and the map distance between genes.

9.  Explain and diagram what is meant by coupling phase and repulsion phase (cis and trans).

10.  Describe how you could go about determining the number of linkage groups for any species,
        and the linkage group for any gene.

11.  If given the map distances between a series of genes obtained from two-point map crosses,
        determine the order of the genes on a chromosome.

12.  Considering three linked genes, and given the numbers of individuals in each phenotype
        which result from a test cross, determine the non-crossover classes, single crossover
        classes and double crossover classes, the position of the gene loci with respect to each
        other, and the map distances apart.

13.  Give a possible explanation of why crossingover seems to occur so precisely between
        genes rather than within genes.

Resources:  Text Chapter 8,  Cartoon Guide pgs. 69-78
 
 

Genetic Linkage and Chromosome Mapping

1903-  Sutton, boveri (cytology + genetics)-  organisms have more "unit factors" than
            chromosomes.

    Soon after ----> discovered that certain genes were not transmitted according to law of
        independent assortment.
        -seemed to be inherited together, transmitted as single unit.

**Organisms have thousands of genes controlling all aspects of its life, therefore, each
    chromosome must carry large numbers of genes (Chromosome theory)

Genes that are part of the same chromosome are said to be Linked and demonstrate Linkage

Chromosomes, not gene, unit of transmission and subject to independent assortment.

        Modifications: Crossing over, provides potential for variation, diversity, survivability

Mendel worked w/ 7 genes, peas have seven chromosomes, he did 21 different crosses.
    Why didn't he find linkage? Lucky??

Bateson, Punnett -----> Coupling

    Example w/ unlinked genes (dihybrid cross)

                        Sepia/ Curved             X              Heterozygous normal
                        sese cc                                         +se +c

    Gametes     se c                                               ++; +c; se+; se c

    Offspring     +se+c                 +se cc                 sese +c                 sese cc
                        normal                 normal curved     sepia normal         sepia curved

                            1:                             1:                         1:                         1

Example w/ linked genes (**Note notation differences**)

        G- grey body     g- black body     R- red     r- purple

P generation             grey/red             X             black/purple

                                    G R                                     g r

                                    g   r                                     g r

                                    (Gg Rr)                                 (gg rr)

Gametes                     G R     g r                             g r

Offspring                     G R                 or                  g r

                                     g r                                         g r

                                    Grey/red                                black/purple                 Expected 1:1 ratio

However!!!!!
Actual results
        black/purple 47%
        grey/ red 47%                         Parental chromosomal types (non recombinant)

        black/ red 3%
        grey/ purple 3%                     Where did these come from? (non parental chromosomal
                                                            types, recombinant)
 

Crossing Over- during meiosis chromosomes replicate forming sister chromatids;
    homologous chromsosmes pair, crossover occurs between non-sister chromatids;
    chromosomes break and rejoin
 
 
 
 
 

from previous example: black/red; grey/ purple resulted from cross overs in heterozygous parent
 
 
 
 

Alleles of douple heterozygotes can appear in two positions:
       Coupling Phase: dominants one one chromosome, recessives on other

                in our example:             G R             Parental non crossovers G R, g r

                                                        g r              Recombinant crossovers G r, g R
                                                                           Least Frequent

       Repulsion Phase: dominant of one locus and recessive allele of other locus on
        same chromosome

            In our example:         G r

                                               g R

        Parental (Non crossovers) G r and g R

        Recombinant (crossovers) G R and g r

    **will make difference in results, parental types are most frequent

    Summary: Recombinant groups due to crossing over
        1. Certain genes assort at random
        2. other genes do not segragate randomly, but are linked, these groups are transmitted
            together
        3. chromosomes occur in pairs (diploids)
        4. linked grnes do not always stay together but are often exchanged (crossing over)
        5. chromosomes are seen to form chiasmata and exchange parts; chiasmata and
            crossover happen with similar frequency.

Using crossover frequencies we can form a linkage map

        Map unit = 1% recombination

        -frequency of crossove is governed by distance between genes

            *as distance increases, frequency of crossover increases

            *short distance between linked genes, fewer crossovers, fewer recombinants

Linkage groups = # of chromosomes in haploid set; Humans, therefore, have 23 linkage groups

Mapping based upon crossover frequencies- Sturtevant
    -studied yellow, white, miniature traits

    -during meiosis limited number of crossovers occur

    -each recombinant event occurs randomly, therefore, the closer two loci reside, less likely
        crossover will cocur between them.

Limits of Recombination
        if 2 genes are located so far apart that the prob. of a crossover occurring is 100%
        then 50% of gemetes will be parental (noncrossover) and 50% will be recombinant
        (crossover). Ex. coin, there is a 100% prob that it will land flat, therefore, we expect a
        1:1 ratio of heads to tails.
 

Genetic Mapping

2 point cross- easiest way to detect crossovers is through test cross progeny

Suppose dihybrid in coupling phase         A C             X             a c

                                                                         a c                              a c

        Progeny        37% AC             37% ac (parental, non crossovers)
                                13% Ac              13% aC (recombinants, crossover)

                           26% of all gametes were crossover types, therefore, distance between
                            A and C = 26 map units
 

Ex. X-linked traits

            Red- w+ (+)             white- w             normal wing- m+ (+)             miniature- m

            P                 w +                             X                     + m
                                w +                                                     Y

                            white eyed normal wing female         red eyed miniature male

F1                           w +                             X                      w +

                                + m                                                     Y
 

F2 (males)             w +              226

                                + m            202                 66.5% parental ( if no crossover you expect 100%)

                                + +              114

                                w m             102                 33.5% recombinant

            ****during meiosis of F1 mother

                    w + --------->                             w m                         + +

                    + m
 

3 Point Test Cross

    Introduction of double crossover groups

    DCO's usually do not occur between genes less than 5 mu's apart. For genes further
    apart it is advisable to use third marker to detect DCO's

    For successful cross:

        1. genotype of organism producing crossover gametes must be heterozygous

        2. phenotype must reflect genotype

        3. large #'s must be produced to recover crossover classes.

        Assumptions:     -parental types (non co's) will be largest group
                                    -double co's will be smallest
 

Trihybrid testcross            A B C                     X                       a b c

                                                a b c                                                 a b c

Progeny                         A B C 36%

                                        a b c 36%                     72% parental, noncrossover types
 

                                        A b c 9%

                                        a B C 9%                      18% single crossovers between A and B
                                                                                (region I)

                                        A B c 4%

                                        a b C 4%                         8% single crossovers between B and C
                                                                                 (region II)

                                        A b C 1%

                                        a B c 1%                          2% double crossovers

To find distance A-B must count all crossovers that occurred in Region I

            CO Region I = 18% + 2% or 20% or 20 map units between A and B

To find distance B-C must count all crossovers that occurred in Region II

            CO Region II = 8% + 2% or 10% or 10 map units between B and C

        Therefore, 30 map units between A and C

Determining Gene Order
        1. Assume any of three possibilities w/ 3 point cross

        2. Observe double crossover groups
            -if you started with coupling phase then only one gene will be changed and
            therefore must be in middle

            Ex. y - w - ec; y - ec - w; w - y - ec

                assume original parent producing crossover was

                    w y ec
                                                if this order correct then DCO's would be w y+ ec, or  w+ y ec+

                    w+ y+ ec+

            After fertilization it was found that the DCO's were w+ y ec and w y+ ec+

            Notice these do not correspond to our predicted

    **Following a double crossover event, allele representing the middle gene will find
        itself present with the outside or flanking alleles present on the opposite parental gamete

        Therefore, our proper order ---->             y+ w+ ec+

                                                                            y   w    ec

Recombinant frequencies aid in determining order
    assume- if co frequencies between a and b is two times greater than between b and c
    then a and b must be twice as far apart on the chromosome that b and c

        Chickens Br (brown eye)        Li (light down)             S (silver plummage)

                10% recombinant freq between Br and Li

                26% recombinant freq between Br and S

                16% recombinant freq between S and Li

            Gene order     Br             Li                             S
 

Ex. Drosophila dumpy wing (dp), clot eye (cl), apterous wing (ap) linked on chromosome #2

            map distances:         dp - ap 42

                                                dp - cl     3

                                                ap - cl   39

                                        dp             cl                                             ap
 
 

Ex. in a testcross                 a b c                 X                    a b c
 
                                               A B C                                      a b c

the rarest classes were abC and ABc

What is the correct sequence?
 

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