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Lecture 16

CAS BI 216 Lecture 16: Genetics - Lecture 16 - Chapter 9 and Chapter 10

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CAS BI 216
John Celenza

Lecture 16 4617 (Chapter 9 and Chapter 10) Chapter 9 cont o Using antibodies to protein A to clone the gene for protein A o Touch filter paper to plate pick up some cells > preserves cell pattern of filter paper lyse cells on filter releases insulin protein put filter into solution w insulin antibody Ab has fluorescent molecule (tag) attached o dry membrane (filter) UV irradiate filter spot (blue) where colony was that contained insulin protein alignment to show where cell is o Cloning by Complementation o YFG+ > white phenyotype cells most cells on plate are YFG+ (white)ura3MATa) YFG => appears black need to isolate and grow a culture of YFG cells transform yeast expression library (collection of vectors with different genes) cDNA library express proteins from yeast from cells that are ura3+ isolate the YFG+ (white) and vectors grow E. Coli sequence insert and characterize YFG+ gene Chapter 10 Genes, Chromosomes, Genomes o DNA Genes Possessed by Different Species o Species o Genome Size (10^6 bp) o Predicted Genes o E. Coli o 4.64 o 4200 o Yeast o 12 o 5800 o Worm o 100 o 19,000 o Fruit Fly o 180 o 13,600 o Mouse o 3000 o 25,000 o Humans o 3000 o 25,000 Fluorescent InSitu Hybridization (FISH) o low resolution assignment of a marker to a chromosome o cells onto plate o slight chromosome nicking (DNase) o Fluorescent DNA probe fro newly identified cloned piece of DNA, which
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