Cloning of bovine integrin β_1 gene and construction of its expression vector

Two pairs of specific primers referring to the bovine integrin beta1 gene in Genebank were designed and synthesized,a 2400 bp fragment was produced by reverse transcription polymerase chain reaction(RT-PCR) from bovine lung.The amplified fragment was cloned to pGEM-T easy vector,confirmed by sequenceing.Then a pair of prokaryotic and euokaryotic expression primers was designed on the basis of sequenceing results.The expression sequences from the cloned vector pGEMbeta1 were amplified and digested respectively by the restricted enzyme EcoR I、Xho I and EcoR I、Xba I.The digested and purified beta1 gene of bovine were cloned into prokaryotic expression vector pET-28a and euokaryotic expression vector pCDNA3.1zero(+).The result showed that the euokaryotic expression recombinant plasmid pCDNAb1 and prokaryotic expression recombinant plasmid pETb1 were successfully constructed.