Position: Home > Articles > RNA interference vector construction and selection of cattle PSMA5 gene and its expression in cow mammary gland epithelial cells
Chinese Journal of Veterinary Science
2016,36
(1)
112-117+131
牛PSMA5基因RNA干扰载体的构建与筛选及在奶牛乳腺上皮细胞中的表达
作 者：
迟玉杨;张晶;徐子娴;高安崇;王秀娜;周长海;牛淑玲;金永成;沈景林
单 位：
吉林大学动物科学学院;吉林大学农业实验基地
关键词：
PSMA5;共轭亚油酸;MAC-T;RNA干扰载体
摘 要：
设计4条待选的shRNA序列以及1条阴性对照序列,构建RNA干扰载体,然后转染到奶牛乳腺上皮细胞(MAC-T)中,最后利用实时荧光定量PCR和Western blotting技术分别检测蛋白酶体α5亚基(PSMA5)在奶牛乳腺上皮细胞(MAC-T)中mRNA和蛋白水平的表达。结果显示:转染shRNA-1组,shRNA-3组和shRNA-4组的PSMA5基因相对表达量显著低于阴性对照组(P<0.05),且shRNA-4组的PSMA5基因相对表达量最低。结果表明:本试验成功构建并筛选出了PSMA5基因RNA干扰载体,并在奶牛乳腺上皮细胞(MAC-T)中成功表达。
译 名：
RNA interference vector construction and selection of cattle PSMA5 gene and its expression in cow mammary gland epithelial cells
作 者：
CHI Yu-yang;ZHANG Jing;XU Zi-xian;GAO An-chong;WANG Xiu-na;ZHOU Chang-hai;NIU Shu-ling;JIN Yong-cheng;SHEN Jing-lin;College of Animal Science,Jilin University;Experimental Base of Agriculture,Jilin University;
关键词：
PSMA5;;conjugated linoleic acid;;MAC-T;;RNA interference vector
摘 要：
This study was to explore the role of proteasome alpha 5subunits(PSMA5)in the period of conjugated linoleic acid(CLA)endogenous synthesis.Firstly,four shRNA sequences and one negative control sequence were designed to construct the RNA interference vector.Then,transfected the RNA interference vector into cow mammary gland epithelial cells(MAC-T).Finally,detected the relative expression of mRNA and protein levels of the PSMA5 gene in the cow mammary gland epithelial cells(MACT)by real-time fluorescent quantitative PCR and Western blotting technology.The results suggested that in transfected shRNA-1,shRNA-3and shRNA-4groups,relative mRNA and protein levels expression of PSMA5 gene was significantly lower than the negative control group(P<0.05).In conclusion,we constructed and selected RNA interference vector of cattle PSMA5 gene successfully,and detected its expression in cow mammary gland epithelial cells(MAC-T).
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