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heterologous gene integration into e.coli genome


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#1 yja97

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Posted 13 February 2013 - 04:48 AM

Hi. I'm trying to integrate heterologous genes into E.coli genome.

The problem is that the size of the genes are about 7 kb.
The only system that I can use is a home-made suicide vector using R6K origin and sacB.

Several trials ended up with failures.
Do you think that 7 kb is too long to integrate into E.coli genome with this system?
Actually, I don't have any experiences to mutate E.coli. Please give me any tips of advice. (incubation time, cautions, troubleshootings etc.)
Thank you in advance.

#2 neuron

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Posted 13 February 2013 - 09:13 PM

I don't know anything about suicide vectors but while searching about this I found out a word document that is attached here. I thought it might be useful for youPosted Image ..

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