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利用PCR技术扩增日本吸血虫凋亡相关基因BAK(Bcl-2 homologous antagonist/killer),并构建重组质粒。应用Realtime PCR技术分析在日本血吸虫非易感宿主大鼠和易感宿主小鼠来源、不同发育阶段虫体中BAK基因的表达状况。结果显示,克隆获得日本血吸虫(Schistosoma japonicum,Sj)BAK编码基因Sj BAK,其ORF含2142bp,编码713个氨基酸,理论分子量为79.3 k Da,理论等电点为4.9。结构域分析表明该基因编码蛋白具有至少3个BH结构域,属于Bcl家族成员。Real-time PCR结果显示该基因在大、小鼠来源4个不同发育阶段虫体均有表达,其中大鼠来源虫体Sj BAK的表达水平都高于小鼠来源虫体,在感染后32 d和42 d表达差异具有显著统计学意义(P<0.05)。由此可推测日本血吸虫凋亡相关基因Sj BAK在血吸虫生长发育中可能发挥重要的作用。
The Bcl-2 homologous antagonist / killer (BAK) gene was amplified by PCR and the recombinant plasmid was constructed. Real-time PCR was used to analyze the expression of BAK gene in different developmental stages of Schistosoma japonicum from non-susceptible host and susceptible host mice. The results showed that the Sj BAK gene encoding Schistosoma japonicum (Sj) BAK was cloned and its ORF was 2142 bp, encoding 713 amino acids. The theoretical molecular weight was 79.3 kDa and the theoretical isoelectric point was 4.9. Domain analysis showed that this gene has at least 3 BH domains and belongs to the Bcl family. Real-time PCR results showed that the gene was expressed in four different developmental stages of large and mouse origin, and the expression level of Sj BAK in the rat-derived parasites was higher than that in the mouse-derived body. After 32 days And 42 d (P <0.05). It can be speculated that Schistosoma japonicum apoptosis-related gene Sj BAK may play an important role in the growth and development of schistosome.