mcb104 handout week 11 - S PATIAL AND TEMPORAL CONTROL OF...

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SPATIAL AND TEMPORAL CONTROL OF GENE EXPRESSION A large fraction of the genome is dedicated to controlling where and when genes are turned on. Genes are turned on and off in specific times/places by molecular enhancers or cis -regulatory elements. Enhancers: -DNA sequences that bind transcription factors and affect transcription of certain genes. -Orientation-independent -Can occur upstream of gene/downstream of gene/in introns/in exons -Usually ~1000 bp away but can be up to 1,000,000 bp away! -DNA sequence conservation is often used to find enhancers You can determine if a conserved sequence is an enhancer by hooking it up to a reporter gene and looking at the expression pattern it drives. Detecting tissue-specific expression Visualizing tissue specific expression: 1) in situ hybridization— for detection of mRNA clone gene make labeled antisense probe hybridize to fixed tissue expressing transcript detect with fluorophore or chemical reaction 2) Immuolocalization— for detection of protein Clone gene make recombinant protein in bacteria purify protein inject into mammalian host isolate antibody specific to protein add protein to fixed tissue detect with secondary antibody Homeosis: A transformation in segmental identity from one fate to another. Example?
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This note was uploaded on 01/28/2010 for the course MCB 104 taught by Professor Urnov during the Fall '09 term at University of California, Berkeley.

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mcb104 handout week 11 - S PATIAL AND TEMPORAL CONTROL OF...

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