How to study the expression of one, specific gene? - (May/21/2006 )

I mean a gene of dehydrogenase in plant. How to write a protocol including a scheme how to study its expression.

I suppose I have to clone specific seqension into vector but which? and how?

Can you help me?

What's wrong with real-time RT-PCR? Just design your own primers and probe set overlapping an exon-exon boundary and you're away. You might even be able to buy preoptimised Taqman Assays.

You don't get much more specific.

I would say that chacking protein expression levels is first, as with western but dont know if applies to plants. Real time PCR is good and then maybe try to overexpress the protein of your interest and see the effects, or siRNA to knock it out
so many things.. so little time

I guess it depends on exactly what you are studying. Saying 'gene expression' is ambiguous. Most people would automatically assume this means the levels of the protein whereas it technically only means the rate at which the gene is transcribed and RNA created.

Do you want to study the protein levels or just if the gene is working?

thanks guys. I've just started studying genetic engeneering so my knowledge is very poor. I want to see if the gene is working only. To see its results.

Well I'd go for real-time PCR in that case. This will tell you if mRNA for your gene is being synthesised.

Can you do this? What platform would you use?