Basic & Clinical Medicine ›› 2011, Vol. 31 ›› Issue (6): 709-712.

Previous Articles     Next Articles

Novel non-viral vector O-CHCS as gene vector for transfecting HBD2 to L929 cells and biological activity detection of expression products

Nan CUI1,Xiao-nan FANG2,Ni ZHANG2,Ming-mao CHEN2,Xue-min LI2,Xin-nian CHEN1   

  1. 1. Lanzhou University
  • Received:2010-07-19 Revised:2010-10-25 Online:2011-06-05 Published:2011-06-06
  • Contact: Xin-nian CHEN

Abstract: Objective To explore O-CHCS’s feasibility of being a gene vector for transfecting L929 cells. Methods We constructed plasmid pCMV- hBD2, and the plasmid was introduced into L929 cells by O-CHCS. Total RNA were extracted from the cultured cells, RT-PCR were performed with specific primers for HBD2 and RT-PCR amplification products were identified with agarose gel electrophoresis;the expression of HBD2 protein was verified by Western blot;the antibacterial activity of purpose protein was defected by Kirby-Bauer disk diffusion method, at the same time put the Lipofect as the positive control. Results RT-PCR amplification products by agarose gel electrophoresis were the same size as the target gene by observation; HBD2 gene expression at the protein level was detected by Western blot ;mediums from the cultured cells transfecting with the pCMV-hBD2 could format antibacterial circle against the Staphylococcus aureus, and these results were as same as the Lipofect. Conclusion The eukaryotic expression vector pCMV-hBD2 was successfully constructed and was introduced to the L929 cells, by doing this we verified O-CHCS could be used as a gene vector for HBD2 , and provided a economic and convenient way to artificial synthesis of HBD2.

CLC Number: