酵母双杂交原理ppt演示.pptVIP

一. 酵母双杂交系统原理(Yeast Two-hybrid System) * * 月饼伯伯 His, β-gal His, β-gal To show you how the yeast-two hybrid system works I must first explain very briefly how gene expression is regulated in Yeast. Here is our gene (in red) ; important in regulating our gene is a so-called UAS where a transcription activator c attach. There are two parts or domains to the transcription activator…the DNA binding domain and the so-called activator domain. If the transcription activator has bound to the the UAS then the activator domain is in a position to interact with the transcription machinery for the gene and bring about gene expression. One last piece of information that is required here is that the transcription activator, being a protein, is also coded in the yeast genome. With this background I now return to our original problem or question…do proteins X and Y bind to each other? In the yeast two-hybrid method we again begin with a yeast gene, this time for a reporter gene, a gene protein the product of which is easy to measure or visualize. This is often the enzyme beta-galactosidase, which if expressed can produces a colored reaction in cells where it comes to expression. * * * To show you how the yeast-two hybrid system works I must first explain very briefly how gene expression is regulated in Yeast. Here is our gene (in red) ; important in regulating our gene is a so-called UAS where a transcription activator c attach. There are two parts or domains to the transcription activator…the DNA binding domain and the so-called activator domain. If the transcription activator has bound to the the UAS then the activator domain is in a position to interact with the transcription machinery for the gene and bring about gene expression. One last piece of information that is required here is that the transcription activator, being a protein, is also coded in the yeast genome. With this background I now return to our original problem or question…do proteins X and Y bind