Cloning promoter (overlap with another gene!) (View forum version)



kkmans

Posted 03 November 2011 - 05:17 AM

Hi everyone,
I have a problem concerning promoter cloning. I find that in my gene of interest [gene A], the 5' upstream region, about 300bp away, is actually another gene [gene B] (transcribed in antisense fashion corresponding to my gene of interest).
In other words, it seems that the antisense strand of the promoter region of gene A is the transcribed region of gene B. Are there any similar case in the literature? Should I just clone the 300bp out? Thanks a lot.

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pDNA

Posted 05 November 2011 - 05:42 AM

looks like a bidirectional promoter?
are we talking about a prokaryotic or eukaryotic system?

Regards,
p

kkmans

Posted 05 November 2011 - 07:38 AM

pDNA,
It is from eukaryotes. So, how I can deal with this strange promoter? Thanks~

pDNA

Posted 05 November 2011 - 02:31 PM

first of all i have to confess i'm no specialist in eukaryotic systems (i like it more simple without a nucleus)
so maybe some specialist can review my advice!

Do you know the exact location of your promoter? ...i would assume it is within those 300 bps or is it further upstream?

just clone the fragment containing your promoter and CDS and shield the bidirectional promoter with an terminator that you clone upstream of your sequence.

Regards,
p