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。
[Objective] To clone Schistosoma Japonicum SJCHGC00821, a novel pairing-assoeiated gene, and to construct its recombinant eukary-otie 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 eukaryotie expression veetor（peDNA3-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 eDNA library of Sehistosoma Japonieum. 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 Sehistosoma Japonicum.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2006年第11期1665-1668,共4页
China Journal of Modern Medicine
基金
国家"十五"重大科技专项课题(No.2002AA2Z3343)
国家863血防重大专项(No.2004AA2Z3530)
湖南省"十五"重点学科建设专项经费联合资助
关键词
日本血吸虫
合抱相关基因
基因克隆
生物信息学
Schistosoma Japonicum
Gene cloning
pairing-associated gene
bioinformatics
