间日疟原虫深圳株红内期SSUrDNA基因片段的克隆与序列分析

目的 体外扩增间日疟原虫深圳株红内期小亚单位核糖体核糖核酸编码基因(SSUrDNA)片段，研究其结构与功能。方法 设计一对特异性引物，采用聚合酶链反应(PCR)从间日疟原虫患者血样中扩增出间日疟原虫SSUrDNA片段，以PUC19质粒T载体构建重组子导入大肠杆菌JM109；阳性克隆双酶切鉴定后，双脱氧末端终止法测定序列。结果 间日疟原虫SSUrDNA扩增片段大小为341bp；阳性克隆双酶切及PCR扩增均得到预期大小的片段；序列测定插入片段为341bp，与Sal I株顺序相比，仅在第151位处缺失一个碱基C。结论 成功克隆了间日疟原虫SSUrDNA片段．该序列在间日疟原虫虫株间高度保守。

Cloning and Sequence Analysis of SSUrDNA Gene Fragment of Plasmodium vivax Shenzhen Isolate

Objective To clone and sequence the blood stage SSUrDNA gene fragment of the Shenzhen isolate of Plasmodium vivax.Methods A pair of primers was designed to amplify the SSUrDNA gene fragment by PCR. The amplified gene fragment was cloned into the PUC19 vector and the DNA sequence of the cloned fragment was determined.Results The SSUrDNA gene fragment, 341bp, of the Shenzhen isolate of P.vivax was amplified and sequenced. A deletion of C at 151bp of the gene fragment was detected.Conclusion The SSUrDNA gene fragment of Shenzhen isolate of P.vivax was successfully cloned and the sequence of the cloned SSUrDNA gene fragment is identical to different P.vivax isolates except for a single base deletion.