isolation of a target gene from a genome

[satokaiba]

Dear all,
Could u suggest me some manners to isolate a target gene from a genome, for instance how can i isolate NOX gene from S.mutans.
Moreover, i would like to know more about gateway system, could u please help me as well
Thanks a lot
Yours scincerely,
satokaiba

[satokaiba]

I wanna support some more informations for my last topic that: I did design the primers for that gene and used NcoI and PstI as the restriction site for it. However, i still wanna have a procedure to do experiments in the lab. Please help:-)
NOX gene is NADH oxidase gene and S.mutans is Streptococcus mutans.

[canalon]

To isolate a gene there are many ways. And it depends what kind of info you have to satrt with.
If you know the sequence, you can design a PCr and clone the PCR product. Or make a restriction of the genomic DNA, clone evrything and look for transformants wich would amplify. That would be a start. If yoou do not know anything about it, well you know the function, there are probably a few sequences of such genes available in gene bank, so use degenerate primers to try to amplify something. Or if you have a phenotypic test, make a knockout of the gene with something like this and insert the plasmid in a recipient strain that is NOX-....

Well want I want to say is that there is not ONE way to reach your goal, and that it depends on what you know. And since I don't now that, I can't be more specific. Sorry