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Lecture

11-29-12.docx

2 Pages
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Department
Biology
Course Code
BIO-0013
Professor
Mc Vey

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11­29­12 • Experiments to show role of enhancers o Run northern blot with complementary probe for gene X o Run another northern blot when enhancer is removed o See difference in the amount of RNA made o Decreased RNA on Northern blot when enhancer is removed • Enhancer experiment o Studied immunoglobulin heave chain/antibody gene found in B cells o B cells make antibodies o Wanted to see if they could take enhancer elements out and still get RNA  for antibodies o Human gene for antibodies  Remove sections of introns, and find one that makes a difference  Use restriction enzyme  Human cell with normal gene—cell 1  Mouse cell without gene inserted—cell 2  Put intact gene in another mouse cell—cell 3  Put modified gene in another mouse cell (deleted intron)—cell4 o When northern blot is done  Cell 1 has high lvls of antibody mRNA  Cell 2 not detectable  Cell 3 high  Cell 4 not detectable • To regulate transcription of a gene, specific regulatory proteins need to bind to the  correct DNA sequences (enhancers, silencers, PPE) • How can you test if enhancers are binding sites for regulatory proteins that  activate transcription? o o Can a B­cell enhancer work on a RBC gene? o B­Cell with just Beta­globin gene o B­Cell with Beta­globin gene and B­cell enhancer o On northern blot  Beta­globin RNA is only det
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