Author(s): XIA Pengpeng, SONG Yujie, DUAN Jinkun, ZHU Guoqiang, Jiangsu Co-Innovation Cen for Important Anim Inf Dis and Zoonoses/Coll of Vet Med, Yangzhou Univ
Pages: 1-4+14
Year: 2013 Issue:  3
Journal: Journal of Yangzhou University(Agricultural and Life Science Edition)
Keyword:  porcine aminopeptidase N (APN); entertoxigenic Escherichia coli (ETEC) F4ac; eukaryotic expression; prokaryotic expression;
Abstract: To investigate the possibility of porcine aminopeptidase N(APN) as a candidate protein receptor for Enterotoxigenic Escherichia coli F4ac,we designed specific primers according to the GenBank sequence on the porcine APN mRNA.The first-strand cDNA was synthesized from total RNA which was extracted from fresh porcine intestinal tissue.Then,we got the whole length of porcine APN gene depending on RT-PCR amplification and constructed both the eukaryotic expression and prokaryotic expression systems respectively.Recombinant plasmid sequencing and restriction enzyme digestion results revealed APN gene had been inserted correctly into both eukaryotic expression vector pcDNA3.1 and pET-28a(+) prokaryotic expression vector,and SDS-PAGE profile showed the recombinant expressing protein was inclusion body proteins with a single clear band of a relative molecular weight of 110KD after purification.The construction of pcDNA3.1-APN and pET28a-TAPN expression systems for Porcine Aminopeptidase N gene and successful expression within the APN protein laid the foundation and platform for further study of porcine APN as a candidate protein receptor for F4ac.