【摘要】 目的克隆与日本血吸虫合抱相关但未知功能的新基因SJCHGC00821,构建其真核表达载体,并应用生物信息学初步探讨其功能。方法应用PCR技术从日本血吸虫成虫cDNA文库中扩增SJCHGC00821基因全长ORF,将其亚克隆到真核表达载体pcDNA3中,通过PCR、限制性酶切分析及测序进行鉴定。应用生物信息学初步分析其细胞定位、蛋白序列、结构域及功能。结果获得日本血吸虫合抱相关未知基因SJCHGC00821的克隆,序列分析结果提示该cDNA序列含有一个597bp的完整阅读框序列,编码198个氨基酸,其编码蛋白的理论分子量为22.76kDa,等电点为4.84。二级结构分析预测提示其具有一定抗原性。结论成功地克隆了日本血吸虫SJCHGC00821基因全长ORF,构建了其pcDNA3真核表达载体,为进一步研究其结构与功能创造了条件。更多还原

【Abstract】 Objective To clone Schistosoma Japonicum SJCHGC00821, a novel pairing-associated gene, and to construct its recombinant eukary-otic expression vector so as to explore its function and roles in pairing. Methods Schistosoma Japonicum SJCHGC00821 full length ORF was amplified by PCR from cDNA library of Schistosoma Japonicum. Recombinant eukaryotic expression vector(pcDNA3-SJCHGC) was then constructed by subcloning technique and confirmed by restriction enzyme digestion analysis and sequencing. Bioinformatic methods were used to analyze its possible structure and function. Results SJCHGC00821, with 597bp base pairs and coding for 198 amino acids, was amplified by PCR from cDNA library of Schistosoma Japonicum. The eukaryotic expression vector corresponding to SJCHGC00821 (pcDNA3-SJCHGC)was successfully constructed,which was confirmed by PCR and sequencing. The molecular weight is 22.76 kDa and the theoretical pI is 4.84. Secondary structure analysis showd that this protein had antigenicity. Conclusions Schistosoma Japonicum SJCHGC00821 gene has been successfully cloned, which will benefit further research on its function as well as its implications in pairing of Schistosoma Japonicum.更多还原