Authors : Jun-Wei Cao, Li-Bing Ma, Yong Zhang, Xiao-Ning He, Cong-Ming Bi, Shu-Ying Peng, Song Hua, Jian-Hong Shu, Wu-JiaFu Sai, Xiang-Chen Li, Chi Zhang and Zhi-Peng Zhang
Abstract: Expression sequences of human Lactoferrin (LF) and Internal Ribozyme Entry Site (IRES) were successfully connected by double enzyme digestion and directed cloning and recombination plasmid pIL was constructed. Mammary gland specific expression plasmid pEBHIL of human lysozyme and human LF was constructed by BamHI restrictive enzyme digestion. Plasmid pEBHIL was introduced to cow mammary gland epithelial cells after liposome transfection. Positive cells were determined by G418 and PCR after EGFP expression observed by fluorescent microscope and proliferated and induced by hormone. The results of Western Blotting analysis on cell supernatant showed that transfected cells had expressed and secreted human lysozyme (MW 14.7 ku) and human LTF (MW 40 ku).
Jun-Wei Cao, Li-Bing Ma, Yong Zhang, Xiao-Ning He, Cong-Ming Bi, Shu-Ying Peng, Song Hua, Jian-Hong Shu, Wu-JiaFu Sai, Xiang-Chen Li, Chi Zhang and Zhi-Peng Zhang, 2012. Coexpression of Exogenous hLYZ/hLF Genes in Bovine Mammary Gland Epithelial Cells. Journal of Animal and Veterinary Advances, 11: 2692-2699.