中国兽医科学2017，Vol.47Issue(12)：1510-1516,7.DOI:10.16656/j.issn.1673-4696.2017.12.007
毒害艾美耳球虫配子体Engam59基因的克隆表达及其表达产物的免疫原性分析
Cloning and expression of Engam59 gene from Eimeria necatrix gametocyte and immunogenicity analysis of its expressed product
摘要
Abstract
Total RNA was isolated from Eimeria necatrix gametocytes.It was used as templates for RT-PCRamplification.A 1 473 bp nucleotide sequence of cDNA had 93.4％ identity to that of Etgam59 of E.tenella.The eDNA ORFencoded a 490-amino acid protein containing a tyrosine-serine-rich region and a proline-glicine-rich region.The fragment of Engam59,aa 211-432,was cloned into the bacterial expression vector and expressed in Escherichia coli BL21 cells.The recombinant gametocyte protein had a molecular weight of 27.69 ku and was predominately expressed in the insoluble inclusion bodies.At last,the specific expression was confirmed by Western-blot using anti-His monoclonal antibody.And it could also be recognized by anti-recombinant protein polyclonal antibodies and convalescent serum.In addition,these findings presented a potential target for future recombinant subunit vaccines against coccidiosis.关键词
毒害艾美耳球虫/Engam59基因/克隆/原核表达/免疫原性分析Key words
Eimeria necatrix/Engam59 gene/cloning/prokaryotic expression/immunogenicity analysis分类
农业科技引用本文复制引用
刘丹丹,曹李琴,朱玉兰,许金俊,陶建平..毒害艾美耳球虫配子体Engam59基因的克隆表达及其表达产物的免疫原性分析[J].中国兽医科学,2017,47(12):1510-1516,7.基金项目
国家重点研发计划项目(2017YFD0501205) （2017YFD0501205）
国家自然科学基金项目(31472181,31602039) （31472181,31602039）
江苏省高校自然科学研究面上项目(15KJB230005) （15KJB230005）
江苏省博士后科研资助计划项目(1601054C) （1601054C）
江苏省高校优势学科建设工程项目 （）
