摘要
【目的】构建能在酵母中表达EBV潜伏膜蛋白 1(LMP1)的穿梭表达质粒 pGBKT7 LMP1。【方法】RT PCR方法扩增EB病毒LMP1全外显子片段 ,利用DNA重组技术将其定向插入细菌 酵母穿梭质粒 pGBKT7的T7启动子下游。【结果】限制性内切酶酶切和DNA测序分析证实RT PCR获得的LMP1cDNA片段与GenBank中的数据完全吻合 ;LMP1准确克隆入 pGBKT7的多克隆位点 ,未改变读码框架。【结论】成功构建了穿梭质粒pGBKT7 LMP1。
To construct bacteria yeast shuttle plasmid pGBKT7 LMP1. The fragment including all the exons of LMP1 was amplified by RT PCR and was recombined to the downstream of T7 promoter in the shuttle vector pGBKT7. It has been proved that the sequence of the RT PCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The LMP1 cDNA fragment was cloned in the vector pGBKT7 in the right direction, and the open reading fragment of LMP1 was maintained. [Conclusion] The pGBKT7 LMP1, a bacteria yeast shuttle plasmid is successfully constructed.
出处
《中山医科大学学报》
CSCD
北大核心
2001年第5期348-351,共4页
Academic Journal of Sun Yat-sen University of Medical Sciences
基金
广东省自然科学基金资助项目 (9940 0 4)
