中国生物工程杂志

Six set primers for influenza A virus polymerase gene PB2、PB1 and PA were designed. With the primers, PB2、PB1 and PA gene were amplified in two parts. Because the primers contain the Ⅱs restriction enzyme cleavage site at 5' end, the amplified products could be cloned into pHW2000 orientionationally post digestion of these fragments with restriction enzymes BsmBⅠ、BsaⅠ or AarⅠ. By detecting the expression of the report gene-enhanced green fluorescent protein (EGFP), the polymerase activity of recombinant plasmids can be proved directly. The cloning strategy was proved to be simple、quick and easy to construct the expression plasmids of influenza A virus polymerases gene for reverse genetics systems, and make latter work easy.