华支睾吸虫SPE16.9基因克隆和序列分析

目的 通过筛选cDNA文库识别华支睾吸虫新基因,为进一步研究其功能及其应用奠定基础。方法 对华支睾吸虫cDNA文库进行筛选,通过Blastn程序进行序列比对,识别华支睾吸虫新基因,并应用软件程序对其进行分析。结果 筛选到一个475bp的核酸序列,编码153aa,理论分子量为16．9kDa,理论等电点（pI）为6．19,疏水氨基酸（AILFWV）占36％,带电荷氨基酸（RKHYCDE）占30％,极性氨基酸（NCQSTY）占28％．酸性氨基酸（DE）和碱性氨基酸（KR）各占9％。经BLAST分析,该蛋白属于ML域（MD-2-related lipid-reeognition）家族．是一个新的分泌蛋白基因。结论 发现华支睾吸虫分泌蛋白基因,可能有作为诊断候选抗原的价值。

Cloning and Sequence Analysis of the Clonorchis sinensis SPE 16.9 Gene

Objective To identify and characterize the new genes of Clonorchis sinensis from a cDNA library. Methods The cDNA library of Clonorchis sinensis was constructed and screened. The cDNA sequences were analyzed with the tblastx and blastn programs on the NCBI website. Results A novel cDNA sequence encoding a secretory protein was identified. Translation of the nucleotide sequence revealed a putative open reading frame of 475bp with a molecular mass of 16.9kDa, pI 6.19 and 36% of "AILFWV", 30% of "RKHYCDE", 28% of "NCQSTY", 9% of each "KR" and "DE". As a result the protein is a putative member of the MD-2-related lipid-recognition family. Conclusion We have identified a novel gene of Clonorchis sinensis encoding a secretory protein, which is a putative member of the MD-2-related lipid-recognition family.