A PiggyBac mediated approach for lactoferricin gene transfer in bovine mammary epithelial stem cells for management of bovine mastitis

// Neelesh Sharma 1, 2, * , Do Luong Huynh 1, * , Sung Woo Kim 3 , Mrinmoy Ghosh 1 , Simrinder Singh Sodhi 1 , Amit Kumar Singh 1 , Nam Eun Kim 1 , Sung Jin Lee 4 , Kafil Hussain 2 , Sung Jong Oh 5 and Dong Kee Jeong 1 1 Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University, Jeju, Republic of Korea 2 Division of Veterinary Medicine, Faculty of Veterinary Science & Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, Jammu, India 3 Animal Genetic Resources Station, National Institute of Animal Science, Rural Development Administration, Namwon, Republic of Korea 4 Department of Animal Biotechnology, College of Animal Bioscience and Technology, Kangwon National University, Chuncheon, Republic of Korea 5 National Institute of Animal Science, Wanju-gun, Republic of Korea * These authors have contributed equally to this work Correspondence to: Dong Kee Jeong, email: newdkjeong@gmail.com , dkjeong@jejunu.ac.kr Keywords: antibacterial milk peptide; bovine lactoferricin; PiggyBac; bovine mastitis; antibacterial activity Received: August 29, 2017      Accepted: September 21, 2017      Published: October 31, 2017 ABSTRACT 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.