Hostname: page-component-8448b6f56d-c4f8m Total loading time: 0 Render date: 2024-04-19T20:14:15.675Z Has data issue: false hasContentIssue false
  • English
  • Français

Purification and antibacterial activity of recombinant human lactoferrin in milk of transgenic mice

Published online by Cambridge University Press:  29 January 2010

Miao Ming-Xing ,
Yuan Yu-Guo ,
An Li-You ,
Zhao Jun-Hui ,
Bai Ya-Jun ,
Guo Lei  and
Cheng Yong
Miao Ming-Xing
Affiliation:
School of Pharmacy, China Pharmaceutical University, Nanjing 210009, China College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
Yuan Yu-Guo
Affiliation:
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
An Li-You
Affiliation:
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
Zhao Jun-Hui
Affiliation:
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
Bai Ya-Jun
Affiliation:
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
Guo Lei
Affiliation:
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
Cheng Yong*
Affiliation:
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
*
*Corresponding author: E-mail: yz.dyx@public.yz.js.cn
Get access Rights & Permissions [Opens in a new window]

Abstract

To verify its antibacterial activity, recombinant human lactoferrin (rhLF) was extracted from the milk of transgenic mice (Mus musculus) (PCL25 and AP) by gel filtration chromatography and analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot and enzyme-linked immunosorbent assay (ELISA). In addition, its bacteriostatic properties were tested using the agar disc diffusion method. ELISA analysis showed that the concentration of rhLF in the milk of transgenic mice ranged from 7 to 8 mg/ml, and the recombinant protein expressed in the milk had the same molecular weight as the native protein (~78 kDa), indicating that the rhLFs had a strong antibacterial activity on Escherichia coli and Salmonella.

Keywords

human lactoferrintransgenic micebacteriostatic properties
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 6 , Issue 3 , December 2009 , pp. 253 - 256
Copyright
Copyright © China Agricultural University 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Waarts, B-L, Aneke, OJC, Smit, JM, et al. (2005) Antiviral activities of human lactoferrin: inhibition of alphavirus interaction with heparin sulfate. Virology 333: 284–292.CrossRefGoogle Scholar
Cao, Y, Gao, HY, Yu, L, et al. (2002) Studies of the clone and cell expression of human lactoferritin gene. Hereditas 24: 9–14 (in Chinese with English abstract).Google Scholar
Chasteen, ND (1983) Transferrin: a perspective. Advances in Inorganic Biochemistry 5: 201–233.Google Scholar
Fujita, K, Ohnishi, T, Sekine, K, Ligo, M and Tsuda, H (2002) Down-regulation of 2-amino-3, 8-dimethylimidazo[4,5-f] quinoxaline (MelQx)-induced CYPIA2 expression is associated with bovine lactoferrin inhibition of MelQx-induced liver and colon carcinogenesis in rats. Japanese Journal of Cancer Research 93: 616–625.CrossRefGoogle Scholar
Tsuda, H, Sekine, K, Fujita, K and Ligo, M (2002) Cancer prevention by bovine lactoferrin and underlying mechanisms — a review of experimental and clinical studies. Biochemistry and Cell Biology 80: 131–136.CrossRefGoogle ScholarPubMed
Huang, HQ and Wang, YZ (2005) Gene clone and Escherichia coli expression of murine lactoferrin. Journal of Agricultural Biotechnology 13: 52–55 (in Chinese with English abstract).Google Scholar
Huang, YZ, Cheng, Y, Bai, YJ and Feng, YJ (2007) Transgenic mice expressing human lactoferrin gene in milk. Progress in Veterinary Medicine 28: 39–43 (in Chinese with English abstract).Google Scholar
Lee, WL, Farmer, JL, Hilty, M and Kim, YB (1998) The protective effects of lactoferrin feeding against endotoxin lethal shock in germ-free piglets. Infection and Immunology 66: 1421–1426.Google Scholar
Mitra, A and Zhang, ZY (1994) Expression of human lactoferrin cDNA in tobacco cells produces antibacterial protein. Plant Physiology 106: 977–981.Google Scholar
Van Berkel, PHC, Welling, MM, Geerts, M, et al. (2002) Large scale production of recombinant human lactoferrin in the milk of transgenic cow. Nature Biotechnology 20: 484–487.CrossRefGoogle Scholar
Farnaud, S and Evans, RW (2003) Lactoferrin, a multifunctional protein with antimicrobial properties. Molecular Immunology 40: 395–405.Google Scholar
Zhang, DB, Zhang, JL, Wang, ZY and Hong, MM (1999a) Expression of cloned human lactoferrin cDNA in transgenic tobacco confers resistance to bacterial and viral diseases. Journal of Plant Physiology and Molecular Biology 25: 234–243 (in Chinese with English abstract).Google Scholar
Zhang, YH, Chi, YJ and Dong, P (1999b) The determination of concentration for lactoferrin by enzyme-linked immunosorbent assay. China Dairy Industry 27: 19–21 (in Chinese with English abstract).Google Scholar
Zhao, YY, Liu, F, Zheng, HY, Lu, Q and Zheng, JG (2006) Expression and bioactivity of human lactoferrin gene in transgenic tomato. Journal of Agricultural Biotechnology 14: 299–300 (in Chinese with English abstract).Google Scholar