逆转录病毒介导的RNAi抑制小鼠生精细胞Zfy基因表达的研究  

A Study of Retrovirus-Mediated RNAi Suppressed the Expressions of Zfy Gene in Spermatogenic Cells of Mice

作　　者：马军德[1] 贾斌[1] 马世俊[1] 申红[1] 张挺军 付维明 蔡国宝 

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]新疆兵团第九师师机关,额敏834609 [3]新疆兵团第九师165团畜牧兽医工作站,额敏834609

出　　处：《石河子大学学报（自然科学版）》2013年第2期176-181,共6页Journal of Shihezi University(Natural Science)

基　　金：新疆兵团博士资金专项(2012BB016);石河子大学研究生创新计划项目(YJCX2010-Y02)

摘　　要：为了构建逆转录病毒RNAi(RNA interference)载体靶向抑制小鼠生精细胞Zfy基因的表达,检测Zfy基因的沉默效果。将构建好的逆转录病毒RNAi载体转染经分离培养的小鼠生精细胞,筛选出稳定转染的单克隆细胞扩大培养,应用qRT-PCR检测各试验组细胞Zfy基因mRNA的表达情况。结果显示,转染逆转录病毒RNAi载体的生精细胞Zfy基因mRNA的表达水平显著降低,与阴性对照和空白对照相比差异极显著(P<0.01);阴性对照与空白对照之间差异不显著(P>0.05)。因此,逆转录病毒RNAi载体可以稳定、高效抑制Zfy基因的表达,为进一步探究Zfy基因在小鼠生精过程中的作用和功能奠定了基础。To construct a novel retroviral vector delivery shRNA suppressing the expressions of Zfy in spermatogenic ceils of mice,and investigate its effect of gene silencing. We transfected the recombinant retroviral vector expression small interfering RNA （siRNA） into spermatogenic cells,and then selected the stable transfected cells scaling up culture. The expression of Zfy gene in cells was assessed by fluorescence quantitation real time PCR （qRT-PCR）. The results showed that the expression of Zfy gene mRNA decreased significantly（P〈0.01）for retroviral vector transfected spermatogenic cells; The expression of Zfy gene mRNA was not statistically significant（P〉0.05）between negatively transfected and untreated cells. In conclusion,retrovi-ral vector could suppress the expressions of Zfy gene effectively and this research lays a foundation for further research into the role and function of Zfy gene in gonepoiesis.

关 键 词：RNAI Zfy基因 逆转录病毒载体 小鼠生精细胞 

分 类 号：S813.3[农业科学—畜牧学] R349.33[农业科学—畜牧兽医]