promoter location

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sahart
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promoter location

Post by sahart » Tue Jan 18, 2011 4:12 am

Greetings,

I am going to clone the promoter of a zebrafish gene. Do you have any idea that how I should find its location?

Thank you :)

Sahar
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canalon
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Post by canalon » Tue Jan 18, 2011 4:25 am

Upstream of your gene.
Sequence from the gene down and see what does have an effect. Or if you have the sequence available, try to amplify and clone the sequence between your gene, and the one before. You can even make multiple slices to identify which part are relevant. Clone in front of gene that express a product easy to track (beta gal is common, but GFP if you can quantify it would be good.
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JackBean
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Post by JackBean » Tue Jan 18, 2011 6:55 am

however, some regulating areas can be even in your gene or downstream of your gene ;)
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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sahart
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Re:

Post by sahart » Wed Jan 19, 2011 3:11 am

canalon wrote:Upstream of your gene.
Sequence from the gene down and see what does have an effect. Or if you have the sequence available, try to amplify and clone the sequence between your gene, and the one before. You can even make multiple slices to identify which part are relevant. Clone in front of gene that express a product easy to track (beta gal is common, but GFP if you can quantify it would be good.


Is it true that if a gene is reverse strand its promoter located at downstream?
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JackBean
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Post by JackBean » Wed Jan 19, 2011 9:00 am

well, it's upstream of your gene, so if you read your gene in reversed complement, than yes, downstream (but there's no need to do it that way. How do you know, which strand is normal and which reversed?)
http://www.biolib.cz/en/main/

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sahart
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Re:

Post by sahart » Wed Jan 19, 2011 9:32 am

JackBean wrote:well, it's upstream of your gene, so if you read your gene in reversed complement, than yes, downstream (but there's no need to do it that way. How do you know, which strand is normal and which reversed?)


well I found this information in ensemble website. it mentioned this gene is reverse strand. What does it mean?
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