基于巢式PCR的重叠延伸PCR优化  

Optimization of Splicing by Overlap Extension PCR Using Nested PCR

作　　者：彭雷[1] 赵艳[2] 马银花[2] 

机构地区：[1]贵州师范大学生命科学学院,贵州贵阳550001 [2]武汉大学生命科学学院,湖北武汉430072

出　　处：《安徽农业科学》2016年第20期126-127,共2页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目"水稻抗褐飞虱基因多样性持续控制褐飞虱的分子机理"(31230060)

摘　　要：[目的]结合巢式PCR对重叠延伸PCR进行优化。[方法]以褐飞虱Actin 1基因启动子区与EGFP表达盒区基因融合为例,通过巢式PCR对重叠延伸PCR进行优化。[结果]成功获得融合片段,经过巢式PCR优化后大大简化试验流程,缩短试验时间,并增强PCR特异性与检出率。[结论]优化了重叠延伸PCR,可更快速高效融合基因片段。[Objective] To optimize the Splicing by overlap extension PCR using nested PCR. [Method] Nested PCR was used to optimize SOE-PCR（ splicing by overlap extension PCR） for combining brown planthopper（ BPH） Actin 1 promoter and EGFP expression cassette.[Result]Nested PCR could greatly optimize SOE-PCR. After optimization of nested PCR,the test process was simplified,and the detection time was shortened. The PCR specificity and detection rate were enhanced. [Conclusion]The optimized SOE-PCR can rapidly and effectively fuse the gene segment.

关 键 词：重叠延伸PCR 巢式PCR 褐飞虱 ACTIN 1启动子 

分 类 号：Q78[生物学—分子生物学]