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#1 zhangyonghe_711

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Posted 09 September 2003 - 07:55 PM

I am going to do clone an enzyme gene on an unknown bacterial genome,I have refered some literatures and found two methods,one is transposon-tagged,the next is useing a certain restriction enzyme to cut genome,so we can get some fragments and then link these fragments to plasmid and transfer them to E.COLI.So I want to get some information about these two methods,and if you have some better methods,can you tell me?thank you.

#2 jonnyr9

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Posted 17 September 2003 - 05:31 AM

Try degenerate PCR, align all known protein sequences for that gene from related bacteria then use CODEHOP to design primers. Carry out PCR with these primers, clone (into pGEM-T) and confim the PCR product identity by sequencing.

There are some releveant links here: http://www.bioinform...520Design.shtml




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