BIOC 300D2 Lecture Notes - Lecture 5: Chemiluminescence, Primary And Secondary Antibodies, Sodium Chloride
8 Jul 2016
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Document Summary
Introduce the concept of rnai based gene knockdown. Understand how to isolate and protect proteins (and 4) introduce immunoblotting as a protein detection method. Interacts specifically with the au-rich elements (ares) in the 3" utr of many mrnas. Is a shuttling protein that carries are-containing mrna between the nucleus and cytoplasm. Hur is a ubiquitous 32 kda protein, contains rna recognition motifs that allow it to bind to mrna. Implicated in many cellular processes including stress response, apoptosis, differentiation, cell-cycle regulation. First evidence in metazoan: anti-sense rna had been used with various degrees of success to reduce protein levels. First demonstration that dsrna is much more potent than anti- sense rna and that successful anti-sense was probably due to dsrna contamination => Function of rnai: (1) as a defense mechanism: many viruses have dsrnas (2) rnai as an endogenous pathway. Rnai actually hijacks an endogenous cellular pathway that produces micrornas (mirnas).
