Yong  Zhang, Jun-Wei  Cao, Li-Bing  Ma, Xiao-Ning  He, Cong-Ming  Bi, Shu-Ying  Peng, Song  Hua, Jian-Hong  Shu, Wu-JiaFu  Sai, Xiang-Chen  Li, Chi  Zhang, Zhi-Peng  Zhang, 
    Coexpression of Exogenous <i>hLYZ/hLF</i> Genes in Bovine Mammary Gland Epithelial Cells,
    Journal of Animal and Veterinary Advances,
    Volume 11,Issue 15,
    2012,
    Pages 2692-2699,
    ISSN 1680-5593,
    javaa.2012.2692.2699,
    (https://makhillpublications.co/view-article.php?doi=javaa.2012.2692.2699)
    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).
    Keywords: Human lysozyme;human lactoferrin;Green Fluorescent Protein (EGFP);transgenic;bovine mammary gland cells