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Gene expression

cloning promoters ORFs

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5 replies to this topic

#1 ott

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Posted 13 June 2012 - 03:46 AM

Hello everyone, i am new in molecular biology and i would like you to help.
i actually want to assess the effect of some chemicals on a cluster of 4 genes (6kb) although two of them seem to be more necessary than others, for the moment i do not know if the best way is to assess the mRNA production or proteins.
second do i need to take out these genes and use a vector for the experiment ?
i have got sequences of my 4 genes from NCBI and i assume ORFs are included and my genes can be expressed.

regards
ott

#2 bob1

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Posted 13 June 2012 - 01:16 PM

a) it depends on what sort of result you want - ideally you should do both.
b)Maybe, is there endogenous expression?

#3 Lastwish

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Posted 17 June 2012 - 09:52 PM

1)both mRNA and protein can be your experimental materials,if one of them can not solve your problem.
2)if you want to do some expression by Prokaryotes,remember cDNA is necessary for your experiment.
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#4 ott

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Posted 21 June 2012 - 08:52 AM

thanks guys i will try both
but i still have a question do we need to make constructs to assess the level of expression?

#5 bob1

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Posted 21 June 2012 - 01:17 PM

For endognous expression, you would look for the expression from the cell only, so no transfected constructs.

#6 ott

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Posted 17 July 2012 - 06:08 AM

Thank you!





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