Mutagenesis and Cloning of HIV-1 gag Gene Fragment by Polymerase Chain Reaction

In the present paper. the primers PG01 and PG02 were synthesized in which the EcoRI and BamHI recognition sites as well as the initiation and termination codons were designed for convenience of subsequent cloning and expression. Micrograms of target gene sequence was produced by 30 rounds of amplification with pJG 423 as a template. The amplified gene fragment was digested with both EeoRI and BamHI and ligated to pUC19 predigested with the same restriction endonucleases. The recombinant plasmid pCY52 was id...