生物信息学方法筛选华支睾吸虫成虫功能基因——钙调神经磷酸酶B亚基样蛋白

目的本文通过建立华支睾吸虫成虫cDNA文库,筛选其功能基因。方法应用SMART方法构建华支睾吸虫成虫cDNA文库,进行大量EST测序,然后应用生物信息学方法将EST序列与GenBank中登陆的序列进行同源性比对、序列拼接及基因完整性判断;并应用NCBI上的RPSBLAST对所筛选基因的保守域进行搜索比对,利用PredictProtein分析、预测其功能域及二级结构。结果从华支睾吸虫成虫cDNA文库中筛选的钙调神经磷酸酶B亚基样蛋白(CsCBLP)基因,经同源性分析,CsCBLP与褐家鼠钙调神经磷酸酶(CaN)同源性为53%,与尾刺耐格里原虫CaNB同源性为40%,与新小杆线虫属结合蛋白的同源性为51%;保守域的比对及功能域、二级结构的预测显示所筛选的CsCBLP是钙调神经磷酸酶B亚基的的类似物,属于钙结合蛋白家族。结论应用生物信息学方法从华支睾吸虫成虫cDNA文库中筛选出钙调神经磷酸酶B亚基样蛋白基因。

Screening of Calcineurin B-like Protein Gene, The Functional Gene of Adult Clonorchis sinensis by Bioinformatical Method

Objective To construct the cDNA library of adult Clonorchis sinensis and screen the fu nctional gene. Methods Construct the cDNA library of adult Clonorchis sinensis by SMART method and sequence a large quantity of EST.Compare the sequence of EST with that reported in GenBank for analysis of homology,then perform sequence splicing and analyze g ene integrity.Analyze the conserved domain of the screened gene by RPSBLAST prog ram in NCBI software and predict its functional domain and secondary structure b y PredictProtein program. Results The calcineurin B-like protein(CsCBLP) gene of adult Clonorchis sinensis wa s screened.The homologies of CsCBLP gene to the CaN of Rattus norvegicus,CaN B of Naegleria gruberi and binding protein of Caenorhabditis elegans we re 53%,40% and 51% respectively.The analysis of conserved domain as well as the prediction of functional domain and secondary structure proved that the screened CsCBLP was analogous to calcineurin B belonging to calcium binding protein fami ly. Conclusion CsCBLP gene was screened from the cDNA library of adult Clonorchis sinensis by bioinformatical method.