How good is PET 23a?

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tkoshy
tkoshy's picture
How good is PET 23a?

Hi All,
I was earlier working with pQE30 vector for cloning. The cloning was fine but I was not getting protein induction, same was the case with other fellow mates who were working with the same vector so now I choose PET 23a for cloning & expression. Can I hav your ideas regarding this? Sequencing, western were done with pQE30 clone to find out the problem & recloning was also tried but all in vain.

Thanks

R Bishop
R Bishop's picture
moved  to cloning  forum.

moved  to cloning  forum. hopefully you get some answers there

Bishop

kjosephs
kjosephs's picture
I only have limited

I only have limited experience with pQE30 but found that I preferred the pET vectors (specifically pET24 and pET28).  pET23 won't be as good as pET24 at suppresssing basal or background expression becauase it lacks a 5 prime lac operator and does not carry the lacI gene.  pET23 also has Amp which is not as good as Kan selection. 

See page 45 in the pET manual linked below.
www.emdbiosciences.com/docs/docs/PROT/TB055.pdf

However, pQE30 is still workable so if you used it correctly and could not detect expression by western blot then you might have a bigger problem.

If you are still trouble-shooting the expression with pQE30 or pET23a maybe you should try writing up what you have done (the more detail you can provide-the better the replys will be) and posting again.