Expression of human β defensin-2 in ectopic loci of endometriosis in rats

Objective To construct pLXSN-hBD2,the human defensin-2 (hBD-2) eukaryotic pLXSN recombinant plasmid,thus allowing for transfection to the ectopic loci of endometriosis in rats (EMS) and subsequent assessment of hBD-2 gene and protein expression.Methods The hBD-2 gene fragment was linked to the digestion sites of EcoR Ⅰ and BamH Ⅰ of the pLXSN plasmid for construction of the recombinant plasmid pLXSN-hBD2 by using double enzyme positioning and directional cloning technology.The EMS rat model was constructed by autologous membrane transplantation.This was followed by multi-foci injection of pLXSN-hBD2 to the ectopic foci,thus allowing for determination of hBD2 protein expression in ectopic endometrium by Western blotting.Results Following recombinant plasmid electrophoresis and sequencing,the designed gene fragment possessed an identical sequence with that of hBD-2 cDNA in GenBank,in which the 54-248 base pairs were consistent with the sequence of hBD-2 target gene.The expression of hBD-2 protein was observed in ectopic foci,as evidenced by Western blotting.Conclusion By multi-foci injection,the recombinant plasmid pLXSN-hBD2 containing hBD-2 gene can be successfully transfected to the endometriosis tissues in rats for harvesting hBD-2 protein. Key words: Defensin-2;  Human;  Recombinant plasmid