巢式RT-PCR分析日本血吸虫组织蛋白酶L1(SjCL1)的基因5'末端序列

目的：测定日本血吸虫组织蛋白酶L1（SjCL1)基因的5'端序列。方法：从日本血吸虫成虫中提取总RNA，以该RNA为模板，进行巢式RT－PCR，扩增SjCL1基因5'端序列。将其与pMD18-T载体连接得到含SjCL1基因5' 端序列的重组质粒，并测定上述DNA插入片段的序列。结果：通过巢式RT－PCR扩增出332bp SjCL1基因5'端序列，测序后，与报道的SjCL1基因部分序列拼接后，可得到一个编码317个氨基酸的完整开放阅读框。结论：使用巢式RT－PCR技术，扩增得到SjCL1基因的5'端序列。为进一步对其进行功能研究奠定了基础。

Sequence analysis of proteinase cathepsin L1(SjCL1)gene 5'-end of Schistosoma japonicum China mainland strain

Aim To analyze the sequence of proteinase cathepsin L1(SjCL1)gene 5' end of Schistosoma japonicum China mainland strain Methods Total RNA was isolated from adult worms of Chinese S japonicum With the RNA as template,the sequence of SjCL1 gene 5' end was obtained by performing nested RT PCR The product of PCR was inserted into pMD18 T vector,and the sequence of inserted DNA fragment was determined Results A 332bp sequence of SjCL1 gene 5' end of Schistosoma japonicum was obtained by using nested RT PCR After combinated with the reported segment of SjCL1 gene,an integrate open reading frame(ORF)was obtained,which encoded 317 amino acids Conclusion The sequence of SjCL1 gene 5' end of Schistosoma japonicum was obtained by using nested RT PCR ,which provided the basis for further study of SjCL1