Dear Eshan,

it depends on where you want to produce the protein. There are different promoters available to express your gene either in bacteria or in eukaryotic cells.

My recommendation is that you amplify the gene from cDNA and ligate your PCR product in a vector called pGem-T. The vector is available as linear DNA with an overhanging T on both ends. For PCRs done with e.g. taq polymerase you always get A overhangs at your products. So you don't have to do blunt end ligations.

Your gene is ligated directly into the lacZ gene and this will give you the possibility to distinguish between empty plasmids and plasmids with insert using IPTG and X-Gal in you agar plates. Just pick some white colonies, prepare some mini preps and check for the right insert by restriction digest.

This vector is commercially available as kit from Promega. I am sure there are several other possibilities from other companies.

For protein expression this pGem-T vector is not feasible because it has no real promoter. This pGem-T vector is just a nice tool for subcloning genes or part of genes and it is rather cheap. But for purification you will have to add a tag (His-tag or whatever) anyway and this needs some further molecular biology.

For protein expression in eukaryotic cells you will need a plasmid with a CMV promoter (e.g. pCDNA3, pTargeT, ...). For expression in E. coli you can take e.g. strain BL21 and use a vector with T7 promoter.

I hope these information will help you.

With kind regards, Oliver

Thank you all for your valuable suggestions.....

check out the Plasmids 101 series of blogs blog.Addgene.org useful tables and info...soon to be an e-book.