Expression of human beta-defensin 3 in COS-7 cell.
Chin Med Sci J
; 19(3): 207-11, 2004 Sep.
Article
in En
| MEDLINE
| ID: mdl-15506650
ABSTRACT
OBJECTIVE:
To establish a cell line for stable expression of human beta-defensin 3 (hBD3).METHODS:
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.RESULTS:
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.CONCLUSION:
We successfully established a hBD3-expressing cell line.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Beta-Defensins
Limits:
Animals
Language:
En
Journal:
Chin Med Sci J
Journal subject:
TERAPIAS COMPLEMENTARES
Year:
2004
Document type:
Article