A PiggyBac mediated approach for lactoferricin gene transfer in bovine mammary epithelial stem cells for management of bovine mastitis
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Neelesh Sharma1,2,*, Do Luong Huynh1,*, Sung Woo Kim3, Mrinmoy Ghosh1, Simrinder Singh Sodhi1, Amit Kumar Singh1, Nam Eun Kim1, Sung Jin Lee4, Kafil Hussain2, Sung Jong Oh5 and Dong Kee Jeong1
1Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University, Jeju, Republic of Korea
2Division of Veterinary Medicine, Faculty of Veterinary Science & Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, Jammu, India
3Animal Genetic Resources Station, National Institute of Animal Science, Rural Development Administration, Namwon, Republic of Korea
4Department of Animal Biotechnology, College of Animal Bioscience and Technology, Kangwon National University, Chuncheon, Republic of Korea
5National Institute of Animal Science, Wanju-gun, Republic of Korea
*These authors have contributed equally to this work
Keywords: antibacterial milk peptide; bovine lactoferricin; PiggyBac; bovine mastitis; antibacterial activity
Received: August 29, 2017 Accepted: September 21, 2017 Published: October 31, 2017
The antibacterial and anti-inflammatory properties of lactoferricin have been ascribed to its ability to sequester essential iron. The objective of the study was to clone bovine lactoferricin (LFcinB) gene into PiggyBac Transposon vector, expression study in the bovine mammary epithelial stem cells (bMESCs) and also to determine the antimicrobial property of recombinant LFcinB against bovine mastitis-causing organisms. The PiggyBac-LFcinB was transfected into bMESCs by electroporation and a three fold of LFcinB secretion was observed in the transfected bMESCs medium by ELISA assay. Furthermore, the assessment of antimicrobial activity against mastitis causing pathogens Staphylococcus aureus and Escherichia coli demonstrated convincing evidence to prove strong antibacterial activity of LFcinB with 14.0±1.0 mm and 18.0±1.5 mm zone of inhibition against both organisms, respectively. The present study provides the convincing evidence to suggest the potential of PiggyBac transposon system to transfer antibacterial peptide into bMESCs or cow mammary gland and also pave the way to use bovine mammary gland as the bioreactors. Simultaneously, it also suggest toward commercial utilization of LFcinB bioreactor system in pharmaceutical industry.
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