15 October 2004

    exam on next Friday, at least 3 exams

    three gene test cross and tetrad analysis not included

     

Funny genetics story and excellent site dealing with dosage compensation - thanks ML !!!!!

                http://post.queensu.ca/~forsdyke/xchromos.htm


Chapter 5 Genetic mapping in eukaryotes


We will not cover the three point test cross or tetrad analysis

This chapter deals with gene linkage and gene mapping. 

The first map was constructed by A. H. Sturtevant while a senior at Columbia University and the favorite student of T. H. Morgan, using the mutations white eye and yellow body.

Reference: Weiner, J. 1999. Time, Love, Memory : A Great Biologist and His Quest for the Origins of Behavior. Knopf New York.

Pictures of Sturtevant (not on web).. ../../Other%20Graphics/Sturtevant1.gif

Example of Gene map, Human Genome Project ... Chromosome22.gif

 

Most important item: learn about parental and recombinant gametes and and parental and recombinant progeny


Series of slides for this...   ../ParentalRecombGams.html


P-generation; the parents and their parental gametes

ParentalRecombGametes1.GIF  

The F-1 and its gametes; which are parental and which are non parental?

We will call the non parental gametes ----- recombinant gametes

ParentalRecombGametes2.GIF

What is the frequency of recombinant gametes?

ParentalRecombGametes3.GIF

Wow! this is independent assortment

           ParentalRecombGametes4.gif



It is best to use a test cross in recombination mapping; why?

        identify parental and recombinant progeny in F-2 ? Dihybrid.gif

 

identify parental and recombinant progeny in test cross ?

ParentalRecombTest1.GIF; 2. ParentalRecombTest2.GIF; 3. ParentalRecombTest3.GIF

Why use a test cross?

The test cross is: a heterozygous individual with one homozygous recessive for the gene or genes in question

like: Ss Yy x ss yy

there is a heterozygous parent and a homozygous recessive parent

In a test cross there is only one genotype for each phenotype; so by looking at the phenotype we may predict the genotype of the gamete from the heterozygote.

In the F-2, with one exception, each phenotype is represented by more than one genotype 

 

Determination of recombination distance between two genes


Series of slides for this .... ../Blackvestigexample.html


Black body and vestigial wing are recessive mutations ...

Recombination1.GIF

Test cross allows for prediction of gametes

eggs? Recombination2a.GIF

eggs filled in Recombination2b.GIF

Test cross data for black and vestigial

The cross

Recombination page3

put in genotypes/sperm

Recombination3a.GIF

predict egg genotypes

Recombination3b.GIF

conclusion: genes are on the same chromosome

Recombination3c.GIF

what is going on?

Recombination page4

from meiosis part of the course

crossingover3.GIF 

conclusions for our two gene example

BlackVestConcl1.GIF

 

The recombination frequency in this cross is 17%

by convention, we say that black and vestigial are 17 map units apart

one percent recombination frequency = one map unit

 1 map unit = 1 centi Morgan (cM)

For a two gene test cross 50% is maximum recombination frequency

one cross over in one cell:

Recombination1OneCell.GIF

one cross over in all cells:

Recombination2AllCells.GIF

one cross over in some cells:

Recombination3OneCell.GIF

Cross over in no cells

Recombination4NoCells.GIF

 

The maximum RF in a two gene test cross is 50%

A RF of 50% is independent assortment; the genes may be on the same chromosome or on different chromosomes.

A RF of less than 50% indicates that the genes are on the same chromosome.

Recombination between genes results from:

the genes being on different chromosomes (inter-chromosomal)

the genes being on the same chromosomes (intra-chromosomal)

***** Random alignment of bivalents in metaphase-I gives inter-chromosomal recomb

***** Crossing over in pachytene gives intra-chromosomal recombination.

In general, the number of cross over events relates to the distance between the genes.

Genes close together on a chromosome will have a low frequency of crossing over

Genes far apart will have a high frequency of crossing over.

Multiple crossover events between two genes will result in an error in calculating map distances.

Two crossovers in all cells

crossingover4.GIF

Again, the maximum recombination frequency in a two gene test cross is 50%

Turns out that frequencies of 10% or lower are accurate; but beyond 10% the observed recombination frequency is an underestimate of the real frequency.

 

Why? Within about 10% there will be only single crossover events; beyond 10% there will be multiple crossovers



18 October 2004

reminder:  Exam on Friday, through chapter 5.  Chapter 21 not included.


And, to finish up chapter 5 ...



Add a third gene cinnabar...

blackvestcbin1.GIF

fit the gene into the map

BlackVestCin2.GIF

 

Concept of linkage groups

As recombination data for a species accumulate genes appear to fall into distinct groups, called linkage groups.

When a large number of genes are mapped the number of linkage groups equals the haploid chromosome number. Surprised at this?

In corn, there are 10 linkage groups and the haploid chromosome number is 10.


Problem of cis/trans (coupling and repulsion in text)

Sequence of slides .....  ../CisTransSequence.html

    Difference between cis and trans..... CisTrans.GIF

    How we get cis..........CisTrcans1.GIF

    How we get trans ......CisTrans2.GIF

    Why the fuss? ..........CisTrans3.GIF


Problem of long distances

    Since the maximum recombination frequency for a two gene cross is 50%, how do we establish distances for genes located a great distance apart?

We add up the short distances of intermediate genes.

LongDistances.GIF


There are several formulae which compensate for long distances. Here are data from one of these:

MappingFunction.gif.... (brief discussion page112)                                                 

Observed RF    Real map units (Kosambi)

        10                          10.1

        30                          34.7

        40                          55.0




Using the recombination map to predict progeny

                    Flash sequence ...../MaptoDataSequence2.html

The problem...................... MapToProgeny1.GIF

The genotypes.................... MapToProgeny2.GIF

Recombination frequency .... MapToProgeny3.GIF

Gamete proportions............ MapToProgeny4.GIF

The answer....................... MapToProgeny5.GIF

Mapping summary ............. MapToProgeny.GI

Examples of genetics maps

    Mendel's seven genes: ../PeaMaps.gif

reference: .....R.C.King, ed. Handbook of Genetics Vol 2

    Also, portion of Drosophila map on practice exam and workbook p.31 

            The Drosophila map:  http://www.ncbi.nlm.nih.gov/books/bv.fcgi?call=bv.View..ShowSection&rid=iga.figgrp.915 



 Complete linkage in male Drosophila melanogaster

CompleteLinkage.GIF 

In humans recombination frequency less in males than in females



early mapping in humans- X-linkage  (considered on first exam)

grandfather method in humans; for certain X chromosome markers for only some pedigrees, not in text

the method:

to determine if a woman is cis or trans

the problem ......................GrandFatherMethod1.GIF

analysis, fill in unknown .....GrandFatherMethod2.GIF

conclusion .......................GrandFatherMethod3.GIF

and again ------ GrandFatherMethod4.gif --- GrandFatherMethod5.gif --- GrandFatherMethod6.gif -- GrandFatherMethod7.gif

early mapping in humans- autosomal

Donahue, R. P., W. B. Bias, and J. H. Renwick. 1968 Probable assignment of the Duffy blood group locus to chromosome 1 in man. PNAS 61: 949-955.

Association with the Duffy blood group with a chromosomal marker

 


Magenis, R. E., F. Hecht, and E. W. Lovrien. 1970 Heritable fragile site on chromosome 16: probable localization of haptoglobin locus in man. Science 170.

Ellen Magenis...magenis.gif

Recombination mapping

Why recombination mapping? A way to organize genes, to learn the biology of recombination, of value for medical counseling.

use of visible markers, like eye color and seed color, in many organisms. Now use of DNA markers.

Example arm lengths for Human chromosomes: http://www.genlink.wustl.edu/info/humgeneral/expal.html

Example of Gene map, Human Genome Project ... Chromosome22.gif

 

Physical mapping

Discusssion in text of physical maps. These are maps which show locations on chromosomes rather than locations via recombination frequencies.

Location of the Xist gene in humans with Flourescent in situ hybridization... xistprob.gif from http://www.pathology.washington.edu/galleries/Cytogallery/cytogallery.html

deletion mapping in Drosophila shows band location of genes, example: DrosMap.gif

 

To chapter 21