Conjugation problems

[Sónia Ferreira]

Dear all,

I have a Klebsiella pneumoniae (KP) and a Citrobacter freundii (CF)
with a 450kb plasmid. In order to see if the plasmid is transmissible
I did a conjugation experiment with KP and CF as donors and E. coli
J53 AZr as recipient. After various attempts I gave up doing it,
because I've never obtained transconjugants. Based in this result is
accurate to assume that the plasmid is non-conjugative?

I also did an electroporation with the same strains as donors and J53
AZr as recipient. I have used a control to make sure the technique was
working. With the control worked, but it didn't work with KP and CF.
Can you give an explanation for this?

best regards

Sonia

[Stephanie Lo]

Dear Sonia,

How can you extract your plasmid? Is the plasmid extraction contain an intact plasmid copy that eligible for transformation? Your plasmid is very large in size.

Steph.

Sónia Ferreira於 2009年10月1日星期四UTC+8上午5時49分40秒寫道：