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具有多个串联重复序列的溶组织内阿米巴膜抗原的克隆与表达。

Cloning and expression of a membrane antigen of Entamoeba histolytica possessing multiple tandem repeats.

作者信息

Stanley S L, Becker A, Kunz-Jenkins C, Foster L, Li E

机构信息

Department of Medicine, Washington University School of Medicine, Saint Louis, MO 63110.

出版信息

Proc Natl Acad Sci U S A. 1990 Jul;87(13):4976-80. doi: 10.1073/pnas.87.13.4976.

Abstract

Entamoeba histolytica causes amebic dysentery and amebic liver abscess, major causes of morbidity and mortality worldwide. We have used differential hybridization screening to isolate an E. histolytica-specific cDNA clone. The cDNA was found to encode a serine-rich E. histolytica protein (SREHP) containing multiple tandem repeats. The structural motif of SREHP resembles some of the repetitive antigens of malarial species, especially the circumsporozoite proteins. A recombinant trpE fusion protein containing the tandem repeats of SREHP was recognized by immune serum from a patient with amebiasis, demonstrating that SREHP is a naturally immunogenic protein. An antiserum raised against the recombinant fusion protein specifically bound to two distinct bands with apparent molecular masses of 46 and 52 kDa in a crude preparation of E. histolytica trophozoite membranes. This antiserum also inhibited E. histolytica trophozoite adhesion to Chinese hamster ovary cells in vitro. The ability to isolate E. histolytica-specific genes, and to express those genes in Escherichia coli, may be important in studying the molecular basis of E. histolytica pathogenesis and for the future development of vaccines.

摘要

溶组织内阿米巴可引发阿米巴痢疾和阿米巴肝脓肿,是全球发病和死亡的主要原因。我们利用差异杂交筛选技术分离出一个溶组织内阿米巴特异性cDNA克隆。发现该cDNA编码一种富含丝氨酸的溶组织内阿米巴蛋白(SREHP),含有多个串联重复序列。SREHP的结构基序类似于疟原虫的一些重复抗原,尤其是环子孢子蛋白。含有SREHP串联重复序列的重组trpE融合蛋白可被一名阿米巴病患者的免疫血清识别,表明SREHP是一种天然免疫原性蛋白。针对重组融合蛋白产生的抗血清在溶组织内阿米巴滋养体膜粗制品中特异性结合到两条明显分子量分别为46 kDa和52 kDa的不同条带上。该抗血清在体外也抑制溶组织内阿米巴滋养体对中国仓鼠卵巢细胞的黏附。分离溶组织内阿米巴特异性基因并在大肠杆菌中表达这些基因的能力,对于研究溶组织内阿米巴发病机制的分子基础以及未来疫苗的开发可能具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/098b/54244/4883bd8e1c00/pnas01038-0092-a.jpg

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