绵羊Zfy基因干扰载体构建及性控效果观察  

Construction of Sheep Zfy gene Interference Vector and Observation of Sexed Control Affection

作　　者：赞哈尔.波拉提 孙丽荣 张永生[1] 贾斌[1] 申红[1] 王绪海[1] 胡政香 

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]内蒙古通辽市农畜产品质量安全中心,通辽028000

出　　处：《石河子大学学报（自然科学版）》2015年第6期721-726,共6页Journal of Shihezi University(Natural Science)

基　　金：新疆兵团科技支疆项目(2014AB012);新疆兵团博士基金项目(2012BB016)

摘　　要：为了研究绵羊Zfy基因干扰载体对性控效果的影响,本试验利用RNA干扰技术针对绵羊Zfy基因m RNA序列设计、构建3个sh RNA干扰载体(p LL3.7-A、p LL3.7-B和p LL3.7-C),并对体外培养的生精细胞进行转染,利用q RT-PCR测定Zfy基因m RNA表达丰度,从而筛选出有效的干扰载体。然后将效果较好的干扰载体通过绵羊睾丸注射,进行体内干扰试验。q RT-PCR检测结果显示:成功构建了3个干扰载体,其中p LL3.7-A、p LL3.7-B对生精细胞Zfy基因的表达水平分别下降了54%(P<0.05)、26%(P<0.05),而p LL3.7-C对其干扰效果不明显(P>0.05);p LL3.7-A、p LL3.7-B干扰载体的体内干扰试验结果显示,后代雌羔率分别为64.7%(P<0.01)、44.4%(P>0.05)。本研究结果表明,本实验构建的干扰载体对绵羊Zfy基因产生干扰作用,且使后代性别比例发生一定程度的偏移,为进一步研究绵羊性别控制提供了理论依据。To research the sheep Zfy gene carrier interference effects on sexual control effect,three recombinant expression vectors were constructed： pLL3.7-A,pLL3.7-B and pLL3.7-C.Then,recombinant expression vectors were transfected to spermatogenic cells in vitro,respectively.Expression level of Zfy mRNA was determined by qRT-PCR.Resuhs showed that pLL3. 7-A and pLL3.7-B significantly inhibited the expression level of Zfy gene,and which were downgrade the expression level of Zfy genes to 54%（P〈0.05）and 26%（P〈0.05）,respectively.And pLL3.7-C had no inhibition （P〉0.05）.Thus,pLL3.7-A and pLL3. 7-B were selected in vivo,and the rate of female offspring of lambs was 64.7% （P〈0.01）and 44.4%（P〈0.05）.It is concluded that the Constrnction of sheep Zfy gene can interfere the vectors and the sex ratio of sheep offspring,which provides a theoretical reference fi^r the further stndy on sheep gender control.

关 键 词：Zfy基因 RNAI 绵羊 性别控制 

分 类 号：S814.1[农业科学—畜牧学] S826[农业科学—畜牧兽医]