Expression of human beta-defensin 3 in COS-7 cell / 中国医学科学杂志(英文版)

ABSTRACT

<p><b>OBJECTIVE</b>To establish a cell line for stable expression of human beta-defensin 3 (hBD3).</p><p><b>METHODS</b>Full length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector identified carrying hBD3 with right direction was introduced into COS-7 cells by Lipofectamine. Cell clones survived in G418-rich medium and with stable expression of hBD3 in both mRNA and protein levels were identified by RT-PCR and Western blot respectively. Genomic integration of the hBD3 gene with the COS-7 cells was confirmed by Southern dot blot and primary analysis. The antimicrobial activity of the secreted hBD3 was also evaluated.</p><p><b>RESULTS</b>COS-7 cells transfected with pcDNA3-hBD3 expressed hBD3 stably in mRNA and protein level. Southern dot blot analysis showed successful integration of the hBD3 gene into the genome of COS-7 cell and the hBD-3 protein secreted into the culture medium showed antimicrobial activity.</p><p><b>CONCLUSION</b>We successfully established a hBD3-expressing cell line.</p>