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杜氏利什曼原虫抗原基因在大肠杆菌中表达的初步研究

[Preliminary study on expression of Leishmania donovani antigen gene in E. coli].

作者信息

Jing B, Hu X

机构信息

Laboratory of Parasitology, West China University of Medical Sciences, Chengdu.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1994;12(2):107-10.

PMID:7955157
Abstract

We constructed previously the expressing library of Leishmania donovani genomic DNA with lambda gt11 as vector. In this paper, 2 x 10(4) phages were plated on E. coli Y1090r-, and screened with a rabbit antiserum prepared by immunization with Leishmania donovani promastigotes. Bound antibodies were detected using alkaline phosphatase labeled anti-rabbit antibodies. A positive expressing clone was detected and transferred into E. coli Y1098r- to prepare lysate, a 39 kDa polypeptide in E. coli Y1089r-lysate was recognized by anti-Leishmania donovani serum. The result indicated that the 39 kDa polypeptide which was not fused with the major portion of beta-galactosidase existed disconnectedly. This finding remained to be further studied.

摘要

我们先前构建了以λgt11为载体的杜氏利什曼原虫基因组DNA表达文库。本文中,将2×10⁴个噬菌体铺在大肠杆菌Y1090r⁻上,并用经杜氏利什曼原虫前鞭毛体免疫制备的兔抗血清进行筛选。使用碱性磷酸酶标记的抗兔抗体检测结合的抗体。检测到一个阳性表达克隆,并将其转入大肠杆菌Y1098r⁻中制备裂解物,杜氏利什曼原虫抗血清可识别大肠杆菌Y1089r⁻裂解物中的一种39 kDa多肽。结果表明,未与β-半乳糖苷酶大部分融合的39 kDa多肽以不连续的形式存在。这一发现有待进一步研究。

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