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[zhangyonghe_711]

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.

[jonnyr9]

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