论文部分内容阅读
目的筛选用于比格犬ERβ基因RNA干扰实验的细胞和检测所用引物。方法根据比格犬ERβ氨基末端区域和DNA结合区设计三对引物,用阳性质粒筛选最佳引物应用于RNA干扰效果检测,并利用293T、Hela和vero细胞的cDNA验证该引物的特异性。对这三种细胞内人ERβ基因的存在情况也进行了检测。结果经过三次重复性实验之后,结果显示引物C36684扩增效率高,且没有非特异条带,该引物对293T、Hela和vero细胞cDNA扩增时同样没有非特异性条带,但293T细胞中存在人的ERβ基因。结论引物C36684用于检测RNA干扰效率,而Hela细胞则作为RNA干扰实验的细胞工具。
Objective To screen the cells for RNA interference experiments on Beagle dog ERβ gene and to detect the primers used. Methods Three pairs of primers were designed according to the amino terminal region of ERβ and the DNA binding region of Beagle dogs. The best primers were screened by positive plasmids for detection of RNA interference and the specificity of the primers was verified by cDNA of 293T, Hela and vero cells. The presence of the human ERβ gene in these three cells was also examined. Results After three repetitive experiments, the results showed that the amplification efficiency of primer C36684 was high and there was no non-specific band. This primer also did not have non-specific bands for cDNA amplification of 293T, Hela and vero cells, but there were human 293T cells Of the ERβ gene. Conclusion Primer C36684 was used to detect RNA interference efficiency, while Hela cells served as a cellular tool for RNA interference experiments.