Promoters and gene expression - (Mar/22/2009 )
Hi, I have a basic question in molecualr biology that I dont understand. When reading some papers for the class I encountered some figures that talk about how certain genes are being expressed using different promoters.
Eg: GFP-SCR driven by a SCR promoter; SHR-GFP driven by a SHR promoter etc. I dont understand what / why you do such experiments using different promoters. By using different promoters what do they expect to see ie: what do they intend to answer, in a general sence.
I dont understand the basic concept of genes being driven by different promoters. Can some one please explain this to me.
Also when fusing SHR gene to GFP what do they expect of it? In general when doing such an experiment.
Some explanations to these would greatly help in understand these concepts and reading the paper. Please help.
thanks!
The promoter used depends on what level of expression you are trying to get and what organism you are trying to express in, some are bacterial only, others are mammalian cell only. Viral promoters give high expression usually.
needhelp on Mar 21 2009, 10:44 PM said:
Eg: GFP-SCR driven by a SCR promoter; SHR-GFP driven by a SHR promoter etc. I dont understand what / why you do such experiments using different promoters. By using different promoters what do they expect to see ie: what do they intend to answer, in a general sence.
I dont understand the basic concept of genes being driven by different promoters. Can some one please explain this to me.
Also when fusing SHR gene to GFP what do they expect of it? In general when doing such an experiment.
Some explanations to these would greatly help in understand these concepts and reading the paper. Please help.
thanks!
GFP is exprssed along with the gene product and as it is fused to it ;by using affinity chromatogrpahy you can purify your gene product form the total amount of proetins because GFP-SCR can stick to the column ,
Lilito on Aug 28 2009, 06:11 AM said:
needhelp on Mar 21 2009, 10:44 PM said:
Eg: GFP-SCR driven by a SCR promoter; SHR-GFP driven by a SHR promoter etc. I dont understand what / why you do such experiments using different promoters. By using different promoters what do they expect to see ie: what do they intend to answer, in a general sence.
I dont understand the basic concept of genes being driven by different promoters. Can some one please explain this to me.
Also when fusing SHR gene to GFP what do they expect of it? In general when doing such an experiment.
Some explanations to these would greatly help in understand these concepts and reading the paper. Please help.
thanks!
In fact, GFP is a green fluorescent protein which is expressed under the control of different promoters, so depending on how powerful is the promoter there is different level of flurescent. And also you can use GFP if you want to see if SHR is expressed and where because it is fused to the protein of SHR so you can detect it.