Expression, purification and identification of recombinant signal peptide of human ?-defensin 2 in A549 cells / 第三军医大学学报

ABSTRACT

Objective To construct high expression system for obtaining human ?-defensin 2 (h?D2) in human lung cancer cell line A549. Methods A recombinant retrovirus expression vector pLNCX2-CEA (signal peptide)-h?D2 (mature peptide) was constructed, and then the retrovirus vector was transfected into PT67 cells by DOTAP. After screening and amplification of single clones by G418, the virus was used to infect A549 cells. A549 cells were also screened by G418, and the cell clones resistant to G418 were obtained. Expression of h?D2 was detected by Western blot analysis. Results A recombinant retrovirus expression vector pLNCX2-CEA-h?D2 was constructed successfully. h?D2 with high level of expression was obtained in A549 cells transfected with the expression vector. Conclusion Human lung cancer cell line A549 can be used for high expression of h?D2 with gene engineering technology.