CSB349L11 - CSB349 Lecture 11 October 20th 2010 Molecular...

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CSB349 Lecture 11 October 20 th , 2010 Molecular genetic and genomic techniques: large scale gene expression analysis ChIP you can do sequencing and do high level of sequences quickly and cheaply and align those sequences to the genome. ChIP is used to identify SS enhancers. They wanted to identify the enhancers responsible for tissue specific. Used it against an anti-body (a HAT), which is found to interact w/ DNA at active enhancers. They chopped up the limbs, forebrain, and midbrain of the embryo and read the genome. They show evolutionary conservation. If the peaks align well, if it really is an enhancer, lets put it inside the embryos and see which expression they’ll get, they see a forebrain expression pattern. Based on enrichment of p300 in forebrain. Drives expression in the limb buds, p300 was pound to the enhancer in thelimb buds, so it’s active there. The binding of p300 only occurred where it was active.
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CSB349 Lecture 11 October 20 th , 2010 We’ve been focused on only DNA, transcriptional control, and RNA transcript. The mechanimsm that regulate genome structure, chromatin modification, mechanisms of transcription (main focus). How information of DNA gets into transcripts. Now we’re going to switch over and focus onto translation. Now we have a full RNA made and need to discuss how it’s regulated.
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CSB349 Lecture 11 October 20 th , 2010 TRANSLATION The binding of sequence specific RNA –binding proteins are more complicated in the sequences they recognise because RNA can take on complex secondary structures. E.g. IRE-BP binds to RNA in presence of iron. There are genes like the feratin H-chain gene. IN the 5’ region of the coding region, a specific structure is formed. IRE structure is formed (regular binding plus weird loops), RNA binding protein recognises only IRE .... inhibits translation initiation by inhibiting the ribosome (leading to reduction of protein expression). The same structure can actually lead to higher protein expression, forms a very complicated structure TfR mRNA recruits IRE which leads to prevention of degredation.
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CSB349
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