Construction and identification of p38 mitogen-activated protein kinases-.BETA. recombinant lentivirus vector carrying small interfering RNA

Objective To construct a p38 mitogen-activated protein kinases (p38MAPK)-β recombinant lentivirus vector carrying small interfering RNA (siRNA) and observe the expression in neuronal cells transfected with this vector.Methods A target-specific sequence from p38MAPK-β mRNA was used to synthesize the corresponding hairpin DNA fragments in vitro.Mter annealing,the DNA products were cloned into siRNA vector to obtain the recombinant siRNA vector plasmid p38MAPK-β-shRNA.The neuronal cells were co-transfected by the lentivirus vector and p38MAPK-β-shRNA,and the recombinant vector of p38MAPK-β-siRNA-lentivirus was obtained.The expression of the transfected genes was evaluated by polymerase chain reaction (PCR) and the titer of purified virus was determined.Results The identification of PCR showed that the recombinant p38MAPK-β-siRNA-lentivirus plasmid was successfully constructed,and the titer of virus was 3 × 10s TU/ml after purification.Conclusion The recombinant lentivirus vector with p38MAPK-β-targeted shRNA is successfully constructed and screened. Key words: p38 mitogen-activated protein kinases-β gene;  RNA interference;  Lentivirus vector