中国兽医科学2017，Vol.47Issue(7)：805-809,5.DOI:10.16656/j.issn.1673-4696.2017.07.001
日本血吸虫胰岛素样多肽的真核表达及其多克隆抗体的制备
Eukaryotic expression and polyclonal antibody preparation of insulin-like peptide of Schistosoma japonicum
摘要
Abstract
In order to clone and express the insulin-like peptide gene(Sjilp) of Schistosoma japonicum and obtain its specific polyclonal antibodies,the primers were designed according to the genome database annotation information,and the cDNA with a length 588 bp from adult worm mRNA was successfully amplified by using RT-PCR.The CDS sequence of Sjilp was 393 bp in length.Sjilp gene codon was optimized based on the codon bias in Pichia pastoris.The recombinant expression vector pPIC9K-Sjilp was constructed and induced in P.pastoris GSll5.Sj-ILP recombinant protein was highly expressed in eukaryotic expression system with a molecular weight of about 15 ku,which was evidenced by SDS-PAGE and Western-blot analysis.Two specific small polypeptides of Sj-ILP were synthesized and linked to KLH,followed by immunization of the New Zealand white rabbits for preparing polyclonal antibodies.The titer of the polyclonal antibodies could reach to 1 ∶51 200 by indirect ELISA analysis.Furthermore,Western-blot analysis showed that the polyclonal antibodies had specifical immunogenicity.This study provided the important foundation for further functional studies between Sj-ILP protein and insulin-like growth factor-binding protein.关键词
日本血吸虫/胰岛素样多肽/真核表达/多克隆抗体Key words
Schistosoma japonicum/insulin-like peptide/eukaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
岳生赟,王帅,何伟,王力,刘仲藜,曾巧英,才学鹏..日本血吸虫胰岛素样多肽的真核表达及其多克隆抗体的制备[J].中国兽医科学,2017,47(7):805-809,5.基金项目
国家重点基础研究发展计划(973)项目(2015CB150300) （973）
