中国兽医科学2017，Vol.47Issue(12)：1486-1491,6.DOI:10.16656/j.issn.1673-4696.2017.12.003
猪带绦虫丝氨酸蛋白酶抑制剂4848的鉴定及表达
Identification and expression of Taenia solium serine protease inhibitor 4848
摘要
Abstract
To identify and express the serine protease inhibitor of Taenia soliam(TsSerpin),the total RNA was extracted from T.solium metacestodes.Specific primers were designed and used to amplify TsSerpin4848 by RT-PCR.PCR products were ligated into the pMD19-T vector.After being transformed into Escherichia coli DH5a,the positive clones were identified and confirmed by sequencing.The sequence of TsSerpin4848 was analyzed with the bioinformatics software.The recombinant pET-30a(+)-4848 plasmid was constructed and transformed into E.coli BL21(DE3).The target protein was expressed,purified and analyzed by SDS-PAGE and Western-blot.In result,TsSerpin4848 cDNA sequence contained an open reading frame of 1 065 nucleotides,and the deduced protein consisted of 355amino acids with the theoretical protein molecular weight of 39 ku.TsSerpin4848 had a serpin conserved structure domain with the DEEGAE and FIVDHPFLFFI,and a specific reactive centre loop (RCL) motif.Analysis of tertiary structure of TsSerpin-4848 revealed that it had eight α-helices and three β-sheets.The expressed products of TsSerpin4848,existed in inclusion bodies,could be recognized by the positive serum from the pig infected with T.solium by immunoblotting,indicating that the protein TsSerpin4848 has a good immunogenicity and it is hopeful to be a diagnostic antigen for cysticercosis.关键词
猪带绦虫/丝氨酸蛋白酶抑制剂/鉴定/表达/候选抗原Key words
Taenia solium/serine protease inhibitor/identification/expression/candidate antigen分类
农业科技引用本文复制引用
梁盼红,刘光学,毛立,张少华,王莉杰,王立群,骆学农..猪带绦虫丝氨酸蛋白酶抑制剂4848的鉴定及表达[J].中国兽医科学,2017,47(12):1486-1491,6.基金项目
国家重点研发计划项目(2017YFD0501303) （2017YFD0501303）
国家自然科学基金面上项目(31372433) （31372433）
