猪肌肉生长抑制素RNA干涉载体的构建与验证  

Construction and Verification of Expression Vector of Myostatin siRNA

作　　者：于永生[1] 罗晓彤[1] 李娜[1] 张树敏[1] 

机构地区：[1]吉林省农业科学院畜牧科学分院,吉林公主岭136100

出　　处：《生物技术》2012年第5期1-5,共5页Biotechnology

基　　金：农业部转基因生物新品种培育项目("抗病转基因猪新品种培育";2008ZX08006-001);吉林省博士后科研启动基金项目("利用转基因技术提高猪生产性能";04005);吉林省农业科学院引进人才基金项目("利用基因转移改善猪肌肉品质";00109)资助

摘　　要：目的:构建针对猪肌肉生长抑制素mRNA的RNA干涉载体,并体内验证其有效性。方法:分别体外转染化学合成4个19 bp的siRNA片段获取针对肌肉生长抑制素的序列,构建RNA干涉载体,将上述载体注射到猪股四头肌中,RT-PCR检测证明其表达有效性及生肌调节因子mRNA水平的变化。结果:获得了针对肌肉生长抑制素的2个小干涉RNA序列,构建了2个重组载体,股四头肌注射混合后的等量重组载体,RT-PCR显示注射重组RNA干涉载体可显著降低肌肉生长抑制素的mRNA水平(约降低了40%),生肌调节因子mRNA水平显著上调(分别约为对照组的2.1～3.9倍)。结论:该试验为在转基因动物的肌肉中表达外源基因奠定了基础。Objective： To construct of RNAi vector which can specifically interfere myostatin transcription and prove its specificity. Method： Four small interfering RNAs （siRNA） target to myostatin were designed and synthesized. After transfection porcine fetal fibroblasts cell by siRNA, the RNA interfering vectors were constructed, and vectors expression specificity were proved by directly injecting the vector into the quadriceps muscle of pig. Result：Two siRNA for silencing of the myostatin gene were achieved and were linked into RNAi vector. The transient expression results indicated that the RNAi vector injection can silence the porcine myostatin gene,in which the MSTN mRNA decreased 40% to the control. The mRNA levels of myogenic regulatory factor markers such as MyoD, myogenin and Myt5 were dramatically elevated 2 -4. 5 times in injection group compared to the control. Conclusion：This study provides the basis for establis- hing transgenic animal which could express porcine myostatin siRNA in the muscle.

关 键 词：猪 肌肉生长抑制素 小干涉RNA 生肌调节因子 体内验证 

分 类 号：Q782[生物学—分子生物学]