山东医药2016，Vol.56Issue(26)：17-19,3.DOI:10.3969/j.issn.1002-266X.2016.26.005
CBS基因过表达PC12细胞株的建立及鉴定
Construction and identification of PC12 ceIIs with overexpressed CBS
摘要
Abstract
Objective To establish stable PC12 cell line with overexpression of cystathionine-β-synthase (CBS)and to identify it.Methods CBS primers were designed according to cDNA sequence of CBS gene from GenBank.The CBS gene was amplified using PCR and ligated into the lentiviral vector GV341.293T cells were transfected with the recombi-nant vector and two kinds of packaging viral vector.Viral supernatant was collected and was used to transfect PC12 cells. After puromycin screening,PC12 cells with CBS gene transfection were constructed.The CBS mRNA and protein of the PC12 cells and the empty vector transfected PC12 cells was detected by real-time fluorescent quantitative PCR and Western blotting,respectively.Results The CBS mRNA expression in the observation group,control group and empty vector group was 162.2 ±11.2,1.0 ±0.03 and 2.3 ±0.07,respectively.The CBS protein expression in the observation group,control group and empty vector group was 1.53 ±0.09,1.00 ±0.10 and 0.96 ±0.10,respectively.The CBS mRNA and protein expression levels of the observation group were higher than those of the control group and empty vector group (P <0.05). The difference of CBS mRNA and protein expression between the control group and empty vector group was not statistically significant.Conclusion The PC12 cell line with overexpressed CBS is successfully constructed.关键词
PC12细胞/胱硫醚-β-合成酶/慢病毒载体Key words
PC12 cells/cystathionine-β-synthase/lentiviral vector分类
生物科学引用本文复制引用
尹蔚兰,廖志强,阳柏成,姜骆永,文芳,邬力祥..CBS基因过表达PC12细胞株的建立及鉴定[J].山东医药,2016,56(26):17-19,3.基金项目
国家自然科学基金资助项目（81200986）；国家级大学生创新创业训练计划项目（201310555196）。 （）
