Construction for retroviral expression vector of high expression and easy isolation of human hBD2

Objective To construct retroviral expression vector of high expression and easy isolation of human hBD2.Methods With designed proper primer, PCR amplifies fusion gene sequence:CEA signal peptide-EK sequence then it is inserted upstream of hBD2 in MCS of pBluescript SK-hBD2 by T4 ligase, with the similar method,thus retroviral expression vector expressing pLNCX2-CEA-EK-hBD2(mature peptide) was constructed. With help of PT67 cell, the CEA-EK-hBD2(mature peptide) is integrated in A549 cell. Results The RT-PCR shows that the A549 cell transcript CEA-EK-hBD2(mature peptide).Conclusion Retroviral expression vector of high expression and easy isolation of human hBD2 was constructed in A549 cell successfully.