期刊文献+

华支睾吸虫19kDa分泌型蛋白(CsSP19)基因的克隆、表达与免疫学功能研究 被引量:1

Cloning,expression and immunological characteristics of the 19 kDa secretory protein from Clonorchis sinensis
下载PDF
导出
摘要 目的对一个新发现的华支睾吸虫分泌型蛋白的未知基因,进行克隆表达和重组产物的免疫学鉴定,确定其免疫学功能。方法利用生物信息学方法从华支睾吸虫成虫全长cDNA质粒文库中发现一个编码分泌型蛋白的新基因(克隆号Cs004f03),将去除信号肽序列的编码序列克隆到原核表达质粒pET-28a(+)中,IPTG诱导表达,产物经亲和层析纯化,并制备大鼠免疫血清,Western-blot方法分析其抗原性和分泌性。结果CsSP19开放阅读框(ORF)有564bp,编码188个氨基酸,理论分子量为21.2kDa,成熟肽的理论分子量为19kDa。该基因的成熟肽编码区在大肠杆菌中可获得高表达,重组蛋白能被华支睾吸虫病人血清所识别,同时,重组蛋白免疫血清也能识别华支睾吸虫分泌排泄抗原中分子量约19kDa的蛋白条带。结论CsSP19蛋白是华支睾吸虫分泌排泄抗原中一个有潜在诊断价值的分泌蛋白。 A cDNA clone encoding a novel secretory protein was isolated from adult Clonorchis sinensis full length cDNA plasmid library.The sequence encoding the mature peptide was cloned into prokaryotic expression vector pET-28a(+)and expressed in E.coli BL21/DE3 after induction with IPTG.Meanwhile,the expressed product after purification with affinity chromatography was used as antigen to prepare rat immune sera.The antigenicity and the secretory property of this protein(CsSP19)were analyzed by Western blotting.It was demonstrated that the open reading frame(ORF)was constituted by 564 bps of nucleotides,encoding 188 amino acids with conceptual molecular weight of 21.2 kDa and the theoretical molecular weight of the mature peptide was 19 kDa.The enoding sequence of the mature peptide could be highly expressed in E.coli.The recombinant protein could be recognized by sera of patients with C.sinensis infection.In addition,sera from rats immunized with the recombinant protein could recognize the secretory protein with molecular weight of 19 kDa in the excretory-secretory antigens of C.sinensis.These results suggest that CsSP19 is a component of excretory-secretory antigens of C.sinensis and may be a potential diagnositic antigen.
出处 《中国人兽共患病学报》 CAS CSCD 北大核心 2007年第11期1079-1082,1087,共5页 Chinese Journal of Zoonoses
基金 "863"专题2006AA02Z422 广东省重大科技专项2004A30801004 广州市科技计划项目2006Z3-E4011
关键词 华支睾吸虫 分泌蛋白 分泌排泄抗原 克隆 表达 Clonorchis sinensis,secretory protein,excretory-secretory antigen,cloning,expression
  • 相关文献

参考文献17

  • 1Lun Zhao-Rong, Robin B Gasser, Lai De-Hua, et al. Clonorchiasis:a key foodborne zoonosis in China[J]. Lancet Infect Dis, 2005,5(1) :31-41.
  • 2Stunell H, Buckley O, Geoghegan T, et al. Recurrent pyogenic cholangitis due to chronic infestation with Clonorchis sinensis [J]. Eur Radiol, 2006,16(11):2612-2614.
  • 3Srivatanakul P, Sriplung H, Deerasamee S. Epidemiology of liver cancer:an over view[J]. Asian Pac J Cancer Prev, 2004,5 (2) : 118-25.
  • 4Nooman Gilani, Mordechai Rabinovitz. Cholangio-carcinoma associated with clonorchis sinensis infection: an unusual cause of biliary tract malignancy [J]. Am J Gastroenterol, 2002, 97 (9):153.
  • 5吴德,余新炳,吴忠道.华支睾吸虫病的流行概况[J].热带医学杂志,2002,2(3):277-279. 被引量:34
  • 6董玉婷,冯正.华支睾吸虫病免疫诊断及分子生物学研究进展[J].国外医学(寄生虫病分册),2003,30(3):108-113. 被引量:5
  • 7Lee Ji-Sook, Kim In Sik, Sohn Woon-Mok, et al. Vaccination with DNA encoding cysteine proteinase confers protective immune response to rats infected with Clonorchis sinensis[J]. Vaccine, 2006,24(3) :2358-2366.
  • 8Lee JY, Kim TY, Gan XX, et al. Use of a recombinant Clonorchis sinensis pore-forming peptide, clonorin, for serological diagnosis of clonorchiasis[J]. Parasitol Int, 2003,52 (6) : 175-178.
  • 9Yang HJ, Park SJ, Kyung-il Im, et al. Identification of a Clonorchis sinensis gene encoding a vitellaria antigenic protein containing repetitive sequences [J]. Mol Biochem Parasitol, 2000,111(9) :213-216.
  • 10张咏莉,吴德,余新炳.华支睾吸虫3-磷酸甘油醛脱氢酶重组蛋白的纯化、酶学活性及免疫学研究[J].中国寄生虫学与寄生虫病杂志,2005,23(4):231-235. 被引量:8

二级参考文献107

共引文献89

同被引文献20

引证文献1

二级引证文献1

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部