Jin Ya-Mei, Lin Jiao-Jiao, Zhang Liang, Fu Zhi-Qiang, Wu Xiang-Fu, Zhou Yuan-Cong, Cai You-Min
Shanghai Institute of Animal Parasitology, Chinese Academy of Agricultural Science, Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Institute for Biological Sciences, Shanghai, China.
Sheng Wu Gong Cheng Xue Bao. 2002 Nov;18(6):698-702.
A 558 bp cDNA fragment was amplified by RT-PCR from adult Schistosoma japonicum(Chinese strain) mRNA with a pair of primers that were designed according to published Sj21.7p gene encoding 21.7 kD protein of Schistosoma japonicum(Philippines strain). Sequence analysis indicated that this frame, named Sj21.7 (Ch), with 99% homology to Sj21.7 p, contained a complete open reading fragment (ORF) of 21.7 kD protein gene of Schistosoma japonicum(Chinese strain). The amino acid sequence shared 98% homology with 21.7 kD protein of Schistosoma japonicum. This fragment was cloned into the expression vector pET28a (+) and subsequently expressed in Escherichia coli with IPTG induction. SDS-PAGE analysis revealed that the molecular weight of this expressed product was 25.4 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.
利用一对根据已发表的编码日本血吸虫(菲律宾株)21.7kD蛋白的Sj21.7p基因设计的引物,通过RT-PCR从日本血吸虫(中国株)成虫mRNA中扩增出一段558bp的cDNA片段。序列分析表明,该框架命名为Sj21.7(Ch),与Sj21.7p具有99%的同源性,包含日本血吸虫(中国株)21.7kD蛋白基因的一个完整开放阅读框(ORF)。其氨基酸序列与日本血吸虫21.7kD蛋白具有98%的同源性。该片段被克隆到表达载体pET28a(+)中,随后在大肠杆菌中用IPTG诱导表达。SDS-PAGE分析显示,该表达产物的分子量为25.4kD。Western印迹表明,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。