日本血吸虫（中国株）雌虫抱雌沟蛋白基因的克隆与表达

[Cloning and expression of gynecophoral canal protein gene of Schistosoma japonicum (Chinese strain)].

作者信息

Jin Y M, Lin J J, Feng X G, Zhang L, Wu X F, Zhou Y C, Cai Y M

机构信息

Shanghai Institute of Animal Parasitology, Chinese Academy of Agricultural Science, Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai 200232, China.

出版信息

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2002 May;34(3):311-7.

PMID:12019443

Abstract

A 1949 bp cDNA fragment was amplified by RT-PCR from adult Schistosoma japonicum (Chinese strain) mRNA with 3 pair of primers that were designed according to published SmGCP gene encoding gynecophoral canal protein of Schistosoma mansoni and SjGCP1 gene encoding the conservative region of gynecophoral canal protein of Schistosoma japonicum. Sequence analysis indicated that this fragment, named SjGCP, with 85% identity to SmGCP, contained a complete open reading frame (ORF) of gynecophoral canal protein gene of Schistosoma japonicum (Chinese strain). The amino acid sequence shared 83.7% identity with gynecophoral canal protein of Schistosoma mansoni. This fragment was cloned into the expression vector pET28c(+) and subsequently expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of this expressed product was 80 kD. Western blotting showed that the recombinant protein reacted well with the rabbit serum immunized with Sj worm antigen, indicating that this expressed product had good antigenicity.

摘要

以日本血吸虫（中国株）成虫mRNA为模板，根据已发表的曼氏血吸虫编码抱雌沟蛋白的SmGCP基因及日本血吸虫编码抱雌沟蛋白保守区的SjGCP1基因设计3对引物，通过RT-PCR扩增得到一条1949 bp的cDNA片段。序列分析表明，该片段命名为SjGCP，与SmGCP的一致性为85%，包含日本血吸虫（中国株）抱雌沟蛋白基因的一个完整开放阅读框（ORF）。其氨基酸序列与曼氏血吸虫抱雌沟蛋白的一致性为83.7%。将该片段克隆到表达载体pET28c(+)中，随后在大肠杆菌中表达。SDS-PAGE显示该表达产物的分子量为80 kD。Western blotting表明，重组蛋白与用日本血吸虫虫体抗原免疫的兔血清反应良好，表明该表达产物具有良好的抗原性。