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Description about the pUC vectors

Description about the pUC vectors - Protocols and Methods Forum

Description about the pUC vectors - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 06-01-2007, 02:57 AM
KiranBabu Tiwari
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Default Description about the pUC vectors



Hi,

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.
The help will be acknowledged.

Thanking you.............

Kiran Babu Tiwari
[Only registered users see links. ]
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  #2  
Old 06-01-2007, 11:51 AM
Aawara Chowdhury
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Default Description about the pUC vectors

In <[Only registered users see links. ].net> ,
KiranBabu Tiwari <[Only registered users see links. ]> wrote:


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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