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目的 寻找日本血吸虫未知基因 ,并对其进行鉴定和编码区克隆。方法 运用表达序列标签 (EST)法寻找日本血吸虫未知基因 ,对获得的表达序列标签运用生物学软件进行分析 ,根据同源基因序列推测其可能的功能 ,并将其克隆入真核表达载体 pEGFP N3中。 结果 获得 1个完整的日本血吸虫cDNA序列 (JAYL0 2 30 ) ,氨基酸同源性分析发现与其他生物的抗凋亡因子 1具有同源性 ,构建了重组克隆 pEGFP N3 JAYL0 2 30。 结论 EST法是一种快速发现和鉴定日本血吸虫未知表达基因的有效方法 ,有助于加速其全长cDNA的克隆。
Objective To search for the unknown gene of Schistosoma japonicum and to identify and clone it. Methods The expressed sequence tag (EST) method was used to search for the unknown gene of Schistosoma japonicum. The expressed sequence tag was analyzed by using biological software. The possible function of the gene was estimated according to homologous gene sequence and cloned into the eukaryotic expression vector pEGFP N3 in. Results A complete cDNA sequence (JAYL0 2 30) of Schistosoma japonicum was obtained. The amino acid homology analysis showed that it has homology with anti-apoptotic factor 1 of other organisms, and the recombinant clone pEGFP N3 JAYL0 2 30 was constructed. Conclusion EST method is an effective method for rapid detection and identification of unknown gene expression in Schistosoma japonicum and is helpful to accelerate the cloning of full-length cDNA.