help: promoter isolation eta

Martin Leach leach at mbcrr.harvard.edu
Sun May 8 21:33:20 EST 1994


In article <2qjo3a$ruf at nermal.cs.uoguelph.ca>, xqiu at uoguelph.ca (Xiao Qiu)
wrote:

> Recently we have isolated a clone from pollen cDNA library of alfalfa which 
> showed significant homology with the alfalfa early nodulin gene. This 
> interesting feature might indicate that the pollen-specific gene is involved
> in recognition and interaction between pollen and stigma, similar with 
> interaction between nitrogen-fixation bacteria and host plants. we plan
> to further charaterize the clone such as test of the hypothesis and its 
> promoter analysis.
> Thus our questions are: 1). how to test our hypothesis that the gene 
> involves cell interaction. 2). is there simple and fast way to isolate
> promoter region only such as PCR or primer extention.
> Any suggestions and help will be greatly appreciated

I have isolated a human neuronal-specific promoter using just PCR
techniques.
Inverse PCR (Triglia et. al N.A.Res), Panhandle PCR, RAGE PCR (rapid
amplification of genomic ends - Biotechniques, 6 months ago), to name but a
few.

Is a bit laborious as in inverse u are guessing at enzyme sites. RAGE PCR
seems the most promising, especially if coupled with the long PCR
techniques (Barnes - may issue of PeNAS) ;)

I posted these refs a month or so ago, if u cant find them email me and i
will post them/mail them

Martin
.
.

-- 

.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
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