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,sequencing and homology analysis of Schistosoma japonicum mitochondrial DNA for large subunit ribosome

目的将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 ,转化大肠杆菌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和用DNA自动测序仪测序。序列送blast基因服务站进行同源性分析。结果构建了3个含日本血吸虫cDNA基因片段的重组子 ,其中1个阳性克隆序列为编码日本血吸虫线粒体大亚基核糖体rRNA的基因序列 ,另2个序列与已知的日本血吸虫基因序列无同源性。结论获得了日本血吸虫线粒体大亚基核糖体rRNA基因片段 ,为进一步分析其一级结构和功能打下了基础。

Aim To determine the gene sequence of positive clones immunoscreened from S.japonicum adult cDNA library, Methods 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 recombinants. The sequences of the inserts were determined using an automated DNA sequencer. Results Recombinants including some S. japonicum cDNA were constructed, one of which encoding S. japonicum mitochondrial DNA for large subunit ribosome. Conclusion Our work provides foundation for further study on the structure and function of S.japoni cum mitochondrial DNA for large subunit ribosome.