A polyclonal antibody against the C subunit of porcine aminopeptidase N expressed in Escherichia coli.
The entire pig aminopeptidase N (pAPN) gene was amplified by RT-PCR using total RNA extracted from intestinal brush border membrane of a newborn piglet. The amplified products of the pAPN gene were cloned into the vector pMD18-T, generating a recombinant plasmid pMD18-T-pAPN. The C subunit of pAPN (pAPN-C) produced by PCR from the plasmid pMD18-T-pAPN was expressed in Escherichia coli using vector pET-32a with His tag. After confirming reactivity of the recombinant protein pAPN-C to antibody against native pAPN, polyclonal antibody against the recombinant protein pAPN-C was prepared in rabbit using purified protein as immunogen. In Western blot analysis, the antibody elicited by the recombinant protein pAPN-C could recognize the native pAPN. These data demonstrate that the pAPN-C recombinant protein and its polyclonal antibody can provide some basis for further receptor antagonist.
Department of Veterinary Clinical Medicine, College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, P.R. China. dongbosun@yahoo.com.cn
Dongbo
D
Sun
Yueqiang
Y
Wang
Guojun
G
Wu
Hong
H
Zhang
Qinghe
Q
Zhu
Xianjing
X
He
Donghua
D
Guo
Rui
R
Wu
D000818:Animals; D000918:Antibody Specificity; D018826:CD13 Antigens / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D003001:Cloning, Molecular; D018352:Coronavirus Infections / Q000517:prevention & control; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D005761:Gastroenteritis, Transmissible, of Swine / Q000517:prevention & control; D005822:Genetic Vectors; D007106:Immune Sera; D016133:Polymerase Chain Reaction; D021122:Protein Subunits / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D013552:Swine
D007106:Immune Sera; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D018826:CD13 Antigens