Description about the pUC vectors

Aawara Chowdhury via methods%40net.bio.net (by aawara from FEMA-trailer.org)
Fri Jun 1 06:51:22 EST 2007


In <mailman.1323.1180696619.5139.methods from net.bio.net>,
 KiranBabu Tiwari <kiranbabu.babukiran from gmail.com> wrote:

> I am working about the cloning and expression of cry proteins from Bt
> (isolated from Khumbu, Mt. Everest region, Nepal) DNNA in E. coli HB 101. I
> should know about the cloning capacity and expression capacity of the
> vectors. Ya, I clonned 4.6kb fragment (of the Bt DNA library) with cry3
> specific fragment amplification using universal primers. I didn't get
> success to express yet. So, please give me more informations about the
> compatibility of the vectors and the host in order to express the cry genes.

Although pUC can be used to express proteins (as a fusion with the alpha
fragment of beta-galactosidase), it is typically not used to express cloned
proteins in E. coli.

We have typically used Qiagen's pQE vector series (express a his-tagged
fusion) in E. coli M15/pDM1.1, or Bill Studier's pET vectors in E. coli
BL21(DE3).  

The pET vectors can also be used in HB101, but you'd have to introduce 
an expression cassette for T7 RNA polymerase into HB101.  This is easily
done by infecting them with lambda CE6.  The pET vectors are sold by
Novagen.

There are a myriad of E. coli strains designed for use with the pET
vectors.

AC
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