猪Zfx基因RNAi载体筛选及性控效果的观察

Construction of RNAi Vecters of Swine Zfx Gene and Their Effects on Sexual Control

为了研究Zfx基因在精子发生过程中的作用,本研究利用RNA干扰技术针对猪Zfx基因m RNA设计3条si RNA并构建sh RNA干扰载体(p LL3.7/e、p LL3.7/f、p LL3.7/g)。对体外培养生精细胞进行干扰后,通过q RT-PCR测定Zfx基因m RNA表达丰度筛选出有效的干扰片段。然后利用有效的干扰载体进行睾丸注射,开展体内干扰试验。结果表明:构建的3个干扰载体,其中p LL3.7/e、p LL3.7/f对生精细胞Zfx基因的干扰率分别为44%(P<0.05)、56%(P<0.05),而p LL3.7/g对其无干扰效果(P>0.05);用p LL3.7/e、p LL3.7/f进行体内试验,结果显示后代仔猪雄性率率分别为51%(P>0.05),59%(P<0.05)。结论:Zfx基因干扰可以影响猪后代的性别比例,本研究可为今后进一步研究动物性别控制提供依据。

To investigate the influence of offspring sex by silencing the Zfx gene during spermatogenesis in boar.Three recombinant expression vectors were constructed：pLL3.7/e,pLL3.7/f and pLL3.7/g.Then,recombinant expression vectors were transfected to spermatogenic cells in vitro,respectively.Expression level of Zfx mRNA was determined by qRT-PCR. Results showed that pLL3. 7/e and pLL3.7/f significantly inhibited the expression level of Zfx gene,and the inhibition rates were 44% （P〈0.05） and 56%（P〈0. 05）,respectively.pLL3.7/g had no inhibition （P〉0.05）.Thus,pLL3.7/e and pLL3.7/f were selected in vivo, and the respectively rate of offspring boar were 51%（P〉0.05） and 59%（P〈0.05）.Zfx gene interference can effectively control the offspring sex of large white, which provides the basis for further research on animal sex control.