TY - JOUR T1 - Coexpression of Exogenous hLYZ/hLF Genes in Bovine Mammary Gland Epithelial Cells AU - Zhang, Yong AU - Cao, Jun-Wei AU - Ma, Li-Bing AU - He, Xiao-Ning AU - Bi, Cong-Ming AU - Peng, Shu-Ying AU - Hua, Song AU - Shu, Jian-Hong AU - Sai, Wu-JiaFu AU - Li, Xiang-Chen AU - Zhang, Chi AU - Zhang, Zhi-Peng JO - Journal of Animal and Veterinary Advances VL - 11 IS - 15 SP - 2692 EP - 2699 PY - 2012 DA - 2001/08/19 SN - 1680-5593 DO - javaa.2012.2692.2699 UR - https://makhillpublications.co/view-article.php?doi=javaa.2012.2692.2699 KW - Human lysozyme KW -human lactoferrin KW -Green Fluorescent Protein (EGFP) KW -transgenic KW -bovine mammary gland cells AB - 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). ER -