you should use BACTOBAC expression system. Topo should be used to clone the genes if you dont have enough from pcr.

is this from Invitrogen? I worked with this system for 3 years, I expressed 4 different proteins but the problem I found was purification and to have a high amount of the recombinant protein for my further analysis where I needed a lot. So the project was never compleated I finished my PhD with a side project that worked. I hope you have more success. Number of experiments I had to optimise from the BACTOBAC manual for it to work e.g. Viral Plaque Assay

good luck

Thanks Merima... but I have another question, The vector pFAST BAC 1, the carrier vector has Amp and Gentamicin resistance and Bacmid present in Bac5alpha competent cell has Tet and Kan resistance, but while antibiotic selection, the manual is saying to plate the transformation mixture on Gentamicin, Tet and Kan resistant plate. Why are they not considering ampicillin resistance? Is it necessary to identify the recombinant colonies by Blue gal???? or there are any other ways???? I have searched for papers and articles but i am not geeting enough information.... can u help me?????

Please see attached protocol for Generating the Recombinant Bacmid

you should use DH10Bac E. coli Competent Cells.

I hope this helps

Merima