Hi..I used pTZ57R/T vector for cloning my gene of interest. Is it necessary to subclone the gene into an expression vector to see the protein expression ? can I induce the protein expression in pTZ57R/T vector itself by using IPTG?
Hi,
Up to my knowledge you can not use pTZ57R/T vector for expression of proteins.
Hi Sandhya,
you will almost certainly need to sub-clone your gene into an expression vector to get expression. Whilst it is not impossible to get expression from pTZ57R/T it would be as a fusion with the LacZ and depending upon orientation and the correct frame.This will probably not be what you want in any case.
To get the best chance for expression an expression vector in an expression strain will be required.
-Richard
Hi Sandhya,
Your gene of interest if it is in frame with LacZ you will have a fusion protein as Richard mention. You for sure to subclone your gene in an expression vector with taking care of all the precautions needed for protein expression such as presence of promoter, ribosomal binding site, ATG etc..;
Abderrahmane
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