日本血吸虫未知功能基因SJFCE3549的克隆及生物信息学分析

Molecular Cloning and Bioinformatic Analysis of a Novel Gene of SJFCE3549

目的克隆与日本血吸虫未知功能的新基因SJFCE3549,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SJFCE3549基因全长ORF,将其亚克隆到真核表达载体pCMV-Tag2A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未知功能基因的克隆SJFCE3549,序列分析结果提示该cDNA序列含有一个453 bp的完整阅读框序列,编码150个氨基酸,其编码蛋白的理论分子量为16.67 kDa,等电点为6.28。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基因SJFCE3549全长ORF,构建了其pCMV-Tag2A真核表达载体,为进一步研究其结构与功能创造了条件。

Objective To clone Schistosoma japonicum SJFCE3549,a novel gene,to construct its recombinant eukaryofic expression vector,and to preliminarily explore its function and roles through bioinformatic methods.Methods The full-length ORF of Schistosoma japonicum SJFCE3549 was amplified by PCR from cDNA library of Schistosoma japonicum.Recombinant eukaryofic expression vector（pCMV-Tag2A-SJFCE3549） was then constructed by sub-cloning technique and confirmed by restriction enzyme digestion analysis and sequencing.Bioinformatic methods were used to analyze its possible structure and function.Results SJFCE3549,with 453bp base pairs and coding for 150 amino acids,was amplified by PCR from cDNA library of Schistosoma japonicum.The eukaryofic expression vector corresponding to SJFCE3549（pCMV-Tag2A-SJFCE3549） was successfully constructed,which was confirmed by PCR and sequencing.The molecular weight was 16.67 kDa and the theoretical pI was 6.28.Secondary structure analysis showed that this protein had antigenicity.Conclusions Schistosoma japonicum gene SJFCE3549 has been successfully cloned,which will benefit further research on its function.