中国动物传染病学报Issue(1)：33-38,6.
日本血吸虫PDIA3的原核表达和多抗血清制备
PROKARYOTIC EXPRESSION OF PDIA3 PROTEIN OF SCHISTOSOMA JAPONICUM AND PREPARATION OF POLYCLONAL ANTIBODIES
摘要
Abstract
The objective of the present study was to express recombinant SjPDIA3 protein in E.coli and prepare polyclonal antibodies. The PDIA3 gene was amplified in PCR using the genome of Schistosoma japonicum as a template. The PCR products obtained were inserted into prokaryotic expression vector pET28a(+). The recombinant plasmid pET28a(+)-SjPDIA3 was transformed into E.coli BL21(DE3). Subsequently, SjPDIA3 protein was expressed with induction of IPTG and purified using NI-NTA resin. The BALB/c mice were immunized with recombinant SjPDIA3 protein to generate polyclonal antibodies. The results showed that a cDNA of 1482 bp was amplified. Bioinformatics analysis determined that the full-length ORF was 1482 bp encoding 493 amino acids and SjPDIA3 protein had a thiol-disulfide oxidoreductase domain and a signal peptide with no transmembrane peptide. Western blot demonstrated prokaryotic expression of the recombinant SjPDIA3 protein with molecular mass of 55 kDa and its efficient antigenicity and immunogenicity as expected.关键词
日本血吸虫/SjPDIA3/原核表达/多克隆抗体Key words
Schistosomajaponicum/SjPDIA3/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
刘群,段明明,苑纯秀,李丹丹,葛程,马丽贞,冯新港..日本血吸虫PDIA3的原核表达和多抗血清制备[J].中国动物传染病学报,2015,(1):33-38,6.基金项目
中央级公益性科研院所基本科研业务费专项(2013JB15)；动物血吸虫病创新团队基金 （2013JB15）
