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Transposon mutagenesis system on Pseudomonas sp.

Transposon mutagenesis system on Pseudomonas sp. - Protocols and Methods Forum

Transposon mutagenesis system on Pseudomonas sp. - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 07-01-2006, 04:45 AM
Jeremy Yuen
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Default Transposon mutagenesis system on Pseudomonas sp.



Hi,

I am a masters student from Malaysia and I'm currently researching on the
genome of Pseudomnas sp. USM 4-55. However, I've problems performing
triparental mating between E. coli DH5 alpha lambda pir (donor strain), E.
coli DH5 alpha (helper strain) with pRK2073 and Pseudomonas USM 4-55.

My questions are as follows

1. What is the best method of performing triparental mating besides filter
mating system?
2. What is the optimised duration of time for mating to occur?


I also conduct biparental mating. But it seems that the plasmid (pUTlacZ1)
could not be transformed into E. coli S 17-1 lambda pir by using TSS.

Can anyone tell me how to transform the plasmid into E. coli S 17-1 lambda
pir?

Thank you.

Regards,

Jeremy
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mutagenesis , pseudomonas , system , transposon


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