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You are here: Home / Associated publications / Belgian Journal of Zoology / Bibliographic References / Construction of expression vector of human lactoferrin and its expression in bovine mammary epithelial cells

Jian Shu, Yong Zhang, Zhi Pan, Shu Peng, Jun Cao and Xiang Li (2007)

Construction of expression vector of human lactoferrin and its expression in bovine mammary epithelial cells

BELGIAN JOURNAL OF ZOOLOGY, 137(2):231-237.

Lactoferrin has multiple biological roles, including inflammation and immune modulation, iron absorption in newborns, non-iron-dependent antimicrobial activity, regulation of the level of retinoblastorna protein, protection against cancer development and metastasis, transactivation of the p53 tumor suppressor gene. Moreover, the ubiquitous expressed human lactoferrin (hLF) is species, tissue, and cell-type specific. To facilitate further studying of the biological function of hLF, we hereby constructed and expressed a recombinant vector containing exogenous hLF gene and bovine mammary gland B-lactoglobulin (BLG) gene. cDNAs of both hLF gene (2259bp) and 5' flank regulatory fragment of BLG (1449bp) consisting of the promoter region, exon 1 and intron 1 of the gene were obtained respectively. Then the translation initiation codon of 5' flank regulatory fragment was mutated from ATG to AAG and renamed as mbIg. hLF and mblg were recombined through PCR method and was named as BL, which was then cloned into pEGFP-Cl vector and renamed pMBL. Later on, the pMBL vector was transferred into a cell line of bovine mammary epithelia by liposomal transfection and cultured with G418 drug for 3 weeks to obtain positive transgene cell clones. Then the expression of hLF was detected in the positive cell clones by Immunocytochemistry and Western Blotting assays. The results showed that the exogenous hLF gene had been successfully integrated into the chromosome of the positive cell clones, which highly expressed hLF.

bovine beta-lactoglobulin (BLG) gene; point mutation; human lactoferrin (hLF) gene; recombinant PCR
BJZ

ISSN 2295-0451 (online version)
ISSN 0777-6279 (printed version)
impact factor 2015: 0,87.

Editor-in-Chief:
Prof. Dr. Isa Schön
Royal Belgian Institute of Natural Sciences
Vautierstraat 29
1000 Brussels, Belgium

 



1863-1903
Annales de la Société malacologique de Belgique
 
1903-1923
​Annales de la Société royale malacologique et zoologique de Belgique
 
1923-1989
Annales de la Société Royale Zoologique de Belgique
 
1989-
Belgian Journal of Zoology