猪Zfx基因RNAi片段的筛选  

The RNAi Fragment Screening of Zfx Gene of Swine

作　　者：郭玉强 程波[2] 宁孟影 贾斌[2] 杜军[3] 戚帅[3] 刘帅兵[2] 王绪海[2] 张永生[2] 韩国华[2] 

机构地区：[1]新疆兵团第九师165团兽医站,额敏县834609 [2]石河子大学动物科技学院,石河子832003 [3]新疆军区农业新技术实验培训基地,石河子830000

出　　处：《石河子大学学报（自然科学版）》2014年第5期550-554,共5页Journal of Shihezi University(Natural Science)

基　　金：新疆兵团博士基金项目(2012BB016);新疆兵团科技支疆项目(2014AB012);石河子大学动植物育种专项(GXJS2013-YZ07)

摘　　要：为了研究Zfx基因在精子发生过程中的作用,本实验利用RNA干扰技术针对猪Zfx基因m RNA的不同靶位设计构建了4个不同的sh RNA干扰载体:3个RNAi-Ready-p SIREN-Retro Q-Zs Green(p SIREN)质粒重组载体(a、b、c),1个p LL3.7慢病毒骨架质粒重组载体(p LL3.7-sh RNA)。通过对仔猪睾丸体外培养生精细胞中的Zfx基因进行人为干扰后,提取总m RNA,利用q RT-PCR对Zfx基因m RNA表达进行丰度测定。结果表明:p SIREN质粒重组载体干扰组对Zfx基因没有明显地作用,慢病毒骨架质粒重组载体干扰组对Zfx基因表达抑制效果显著,抑制率为51%(P﹤0.05)。结论:筛选获得了慢病毒骨架质粒重组干扰载体,其对仔猪睾丸体外培养生精细胞中的Zfx基因表达具有显著地干扰作用。The research was aimed at studying the role of Zfx gene in the process of spermatogenesis.Four different shRNA interference vectors were designed and constructed by RNA interference for the mRNA of swine Zfx in this study, including three RNAi-Ready-pSIREN-RetroQ-ZsGreen （pSIREN） plasmid vectors and a Lentivirus skeleton plasmid vector. After interfering with the expression of spermatogenic Zfx Gene of Swine testis in vitro, the total mRNA was extracted from spermatogenic cells,and abundance determination of the Zfx gene mRNA level was analyzed by qRT-PCR.The results showed that the three pSIREN plasmid vectors had no significant effect on inhibiting Zfx gene,while the lentivirus plasmid effectively inhibited the expression of the Zfx gene the inhibition rate was 51%.In conclusion,we successfully obtained a Lentivirus skeleton plasmid vector which could inhibit the expression of spermatogenic Zfx Gene of Swine in vitro.

关 键 词：ZFX RNAI 猪 性别控制 

分 类 号：S813.3[农业科学—畜牧学]