BIOL 430 2010 lecture 7 handout

BIOL 430 2010 lecture 7 handout - BIOL 430 Lecture BIOL 430...

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IOL 430 Lecture 7 BIOL 430 Lecture 7 January 26 th 2010 ow do we identify genes involved in development? How do we identify genes involved in development? Making transgenic animals Mosaic analysis BIOL 430 2010 Molecular Genetics of Development Dr. Ian Chin-Sang www.chin-sang.ca
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How do we Isolate Genes involved in Development? Mutagens Type of Type of DNA Applications Mutagen Damage Chemical mutagenesis(i.e. EMS or Base Pair Change point mutation Saturation mutagenesis, fficient (1/1000) ENU N-ethylinitrosourea (ENU) ehtylmethanesulfonate MS) GC to AT efficient (1/1000), random, isolate any type of mutation lof or gof (EMS) Transposons (i.e. P element or Tc1) DNA insertion not quite random, provides a “tag” for loning isolate loss of cloning, isolate loss of function mutations as well as gain of function mutations BIOL 430 2010 Molecular Genetics of Development Dr. Ian Chin-Sang www.chin-sang.ca Irradiation (X rays γ rays) Deletions and chromosomal rearrangements inefficient (1/10 000) random, usually isolate null mutations
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Identification of recessive Mutations n zebra fish Male fish are mutagenized (ENU) rare event leads to mutation m/+ 1000’s of individual crosses required m/+ +/+ Raise families 50% are +/+ 0% are =+/m m/+ +/+ m/+ +/+ or or 50% are +/m Only the double het cross will ive rise to 25% mutant progeny m/+ m/+ m/+ m/+ +/+ +/+ +/+ +/+ give rise to 25% mutant progeny BIOL 430 2010 Molecular Genetics of Development Dr. Ian Chin-Sang www.chin-sang.ca 25% of progeny mutant can maintain as homozygote or as a het if the mutation is lethal BOX 3B p. 99
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a b c d e f X ene “c” is recessive lethal mutation a b c d e f X 5’ 3’ gene c is recessive lethal mutation Dead (homozygous m/m) 5’ 3’ a b c d e f gene “c” is recessive lethal mutation a b c d e f X BIOL 430 2010 Molecular Genetics of Development Dr. Ian Chin-Sang www.chin-sang.ca Alive (heterozygous +/m)
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a b c d e f a b c d e f cross gene “c” is recessive lethal mutation a b c d e f 5 3 a b c d e f 5 3 Alive (heterozygous) Alive (heterozygous) a b c d e f 5% alive +/+ a b c d e f 25% alive +/+ Phenotypically theses look the same. a
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This note was uploaded on 12/06/2011 for the course BIOL 430 taught by Professor Dr.ianchin-sang during the Winter '10 term at Queens University.

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BIOL 430 2010 lecture 7 handout - BIOL 430 Lecture BIOL 430...

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