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[Expression of alpha-defensin-1 with J chain in the transfected COS-7 cells].

OBJECTIVE
To recombine the alpha-defensin-1 (HNP-1) and J chain cDNA fragment into a new germicidal molecule J-HNP-1, and investigate its expression in the transfected COS-7 cells.


METHODS
J chain and HNP-1 cDNA were amplified from the plasmids pCH-J and pBabeNeo-HNP-1 respectively, then the J-HNP-1 DNA fragment was obtained by recombinant polymerase chain reaction (PCR). The J-HNP-1 fragments were inserted into pcDNA3.1 (-)/Myc-HisC, which subsequently was made to transfect the COS-7 cell by lipid transfection method. The expression was confirmed by Western blot.


RESULTS
Determination with His antibody, a dense band at 24 ku was displayed by the cell lysate and the cell culture supernatant in concordance with that of the predicted extent, but not in the control.


CONCLUSION
The new germicidal peptide J-HNP-1 is found to be expressed intracellularly and could be secreted. The results lay the foundation for further study on the germicidal activity of J-HNP-1.