Expression and functional analysis of porcine aminopeptidase N produced in prokaryotic expression system.
Porcine aminopeptidase N (pAPN) is a cellular membrane protein and a functional receptor for porcine coronaviruses. Here, we describe the heterologous expression of pAPN without signal peptide in BL21(DE3)pLysS host cells. The Escherichia coli (E. coli) harboring the recombinant construct was efficiently induced to express the pAPN protein at a high level. The most optimal expression profile for pAPN expression was investigated. By inoculating a rabbit with the purified pAPN, a high tittered specific antibody was achieved. Biologically, the antibody reacted with either pAPN-expressing E. coli or native pAPN on the surface of swine testis cells. The pAPN and its specific antibody blocked transmissible gastroenteritis coronavirus infection in vitro. Furthermore, the localization of pAPN on the small intestine of swine was analyzed by immunohistochemistry.
Liu
Boqi
B
College of Veterinary Medicine, Northeast Agricultural University, Xiangfang District, Harbin, China.
Li
Guangxing
G
Sui
Xiuwen
X
Yin
Jiechao
J
Wang
Heng
H
Ren
Xiaofeng
X
D000818:Animals; D018826:CD13 Antigens / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D017934:Coronavirus / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D007150:Immunohistochemistry; D008297:Male; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D013552:Swine
D011994:Recombinant Proteins; D018826:CD13 Antigens