中国兽医科学2017，Vol.47Issue(3)：375-380,6.DOI:10.16656/j.issn.1673-4696.2017.03.016
猪ISG15基因的克隆表达与多克隆抗体的制备
Cloning and expression of pig ISG15 gene and preparation of the polyclonal antibodies
摘要
Abstract
The ISG15 gene of pig was amplified by RT-PCR from peripheral blood lymphocytes cells.The pig ISG15 gene was cloned into pET28a,expressed with a 6×histidine tag by IPTG and purified by Ni2+ affinity chromatography.Rabbits were immunized with purified ISG15 protein to obtain the polyclonal antibodies.The pig ISG15 gene was found to consist of 504 bp encoding 167 amino acid residues.Compared with ISG15 gene of other species,the most conserved regions of the pig ISG15 peptide were found to be the tandem ubiquitin-like domains (No.3-75 amino acids in theN-terminal and No.81-156amino acids in the C-terminal)and LRLRGG conjugating motif in the C-terminal,characteristic of the ISG15 proteins.Western-blot analysis showed that the antibodies had good specificity.The ELISA results showed that the antibody titer of polyclonal antibodies was 1∶102 400.This study result lays a foundation for further studies of the function of pig ISG15.关键词
猪/ISG15基因/蛋白表达/多克隆抗体Key words
pig/ISG15 gene/expression/polyclonal antibodies分类
农业科技引用本文复制引用
李双双,李新果,石昂,时庆贺,杨利,王玉国,陈陆,李永涛,王川庆..猪ISG15基因的克隆表达与多克隆抗体的制备[J].中国兽医科学,2017,47(3):375-380,6.基金项目
河南省高校科技创新人才支持计划项目(14HASTIT022) （14HASTIT022）
国家自然科学基金资助项目(31272567) （31272567）
河南省高校科技创新团队支持计划资助项目(14IRTSTHN015) （14IRTSTHN015）
