1.Molecular Cloning of Two Novel Temporins From Lithobates catesbeianus and Studying of Their Antimicrobial Mechanisms
Ruili ZHAO ; Junyou HAN ; Wenyu HAN ; Liancheng LEI ; Changjiang SUN ; Xin FENG ; Lina JIANG ; Hongwei QIAO ; Linjun CAI
Progress in Biochemistry and Biophysics 2009;36(8):1064-1070
Temporins are a kind of small,hydrophobic and C-terminus amidated antimicrobial peptides from Rana species.They are effective against bacteria,fungi,yeast,protozoa and viruses.Two novel temporins named as temporin-La(LLRHWKILEKYLanifc) and temporin-Lb(LFRHVVKIFEK.Ylamid.) were cloned from Lithobates catesbeianus.Synthetic peptides of temporin-La and temporin-Lb showed strong antimicrobial activities against bacteria tested,especially Gram-positive bacteria.Besides,temporin-La showed no haemolytic activity to rabbit erythrocytes at the concentration of 250 mg/L while temporin-Lb showed weak haemolytic activity(LC50≈ 230 μmol/L).Transmission electron microscopy showed that temporin-La and temporin-Lb induced different effects on bacterial structure of Staphylococcus aureus.
2.Expressing of bovine viral diarrhea-mucosal disease virus Changchun184 strain of E2 gene in BCG
Rui DU ; Yan DIAO ; Junyou HAN ; Xichen ZHANG ; Kun SHI
Chinese Journal of Veterinary Science 2009;29(7):854-857
E2 gene of BVDV Changchun 184 strain was cloned and inserted into the shuttle expression plasmid vector pMV261,the recombinant shuttle plasmid pMV261-E2 was constructed.Then pMV261-E2 was transformed into BCG successfully and obtained recombinant BCG which was resistive to kanamyein.The recombinant BCG were identifieated by PCR.E2 gene expression in recombinant BCG was induced in 45℃,then the SDS-PAGE and western blotting was used to analyze the expression product.The results indicated the BVDV E2 gene was expressd in BCG successfully.

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