编码日本血吸虫线粒体大亚基核糖体基因的亚克隆、测序及同源性分析

CLONING AND SEQUENCING OF THE GENE ENCODING THE SCHISTOSOMA JAPONICUM MITOCHONDRIAL DNA FOR LARGE SUBUNIT RIBOSOME

目的筛选日本血吸虫成虫 cDNA 文库,得到基因克隆并测序。方法体外将以阳性克隆为模板的 PCR 产物和 pGEM-T 载体连接,转染大肠杆菌 XL1-blue,经抗生素及生色底物 X-gal 初筛,再用限制性内切酶酶切法进一步鉴定为重组质粒后,DNA 自动测序仪测序。序列送blast 基因服务站进行同源性分析。结果构建三个含日本血吸虫 cDNA 基因片段的重组子,其中一个阳性克隆序列为编码日本血吸虫线粒体大亚基核糖体的基因序列。结论获得编码日本血吸虫线粒体大亚基核糖基因片段,为分析其作为候选重组疫苗分子的潜能打下基础。

Aim To determine the gene sequence of positive clones immunosereened from S.japonicurn adult cDNA li- brary,Method The products of PCR were ligated into pGEM-T vector and then used to transform E.coli XL1 -blue.Following blue and white selection screening,two restriction enzymes were used to characterize the re- combinants.The sequences of the inserts were determined using an automated DNA sequencer.Results Re- combinants including some S.japonicurn cDNA were constructed,ond of which encoding S.japonicum mito- chondrial DNA for large subunit ribosome.Conclusion Our work provides foundation for vaccine trial probably using S.japonicum mitochondrial DNA for large subunit ribosome.