Hello everyone,
since my supervisor left already I am desperately looking for the restriction map of plasmid pMHH1. I want to start curing my E. coli of its plasmid by using suicide vector pMHH1. In order to start cloning my PCR product I need to know the restriction sites of pMHH1 and can´t find it anywhere? Please help me out!
Hi Stefan,
Is it pMH1 vector? Restriction map is attached. Good luck!
Hi Ying,
I am afraid it really is pMHH1. Suppose this is a rare suicide vector although it has been used by Coldewey SM, Hartmann M, Schmidt DS, Engelking U, Ukena SN, Gunzer F - (2007) already. Maybe you can find. Thanks in advance!
From Figure 3, looks like SacI and XbaI will make single cut in vector. Can you use these sites?
Xba I definitely. But I need to know whether I can use Xma I since my insert has both restriction sites cloned?
Check with sequence of pGP704, there is a Xma I site just 30 bp downstream of XbaI. Since pGP704 is only 3.7 kb and pMHH1 is 5.6 kb. I don't know if the newly added sacB has the XmaI site(s) in it.
Checked sacB sequence (1.9 kb), no Xma I was detected. Good luck in cloning!
Maybe a good alternative to pMHH1 but I cannot use pGP704 as suicide vector, can I ?
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