响应重力负荷变化的抗肌萎缩表达载体构建  

Construction of Anti Muscular Atrophy Expression Vector under the Control of Loading Sensitive Promotor

作　　者：车速 黄庆生[1] 李琦[1] 叶琳洁[1] 杨慧[1] 

机构地区：[1]西北工业大学/生命学院/空间生物实验模拟技术国防重点学科实验室,陕西西安710072

出　　处：《生物技术》2012年第6期23-27,共5页Biotechnology

基　　金：国家自然科学基金项目(30971425)资助

摘　　要：目的:为了验证‘肌肉生长抑制素’启动子(PM)对去负荷(失重)的响应能力,观察PM调控下的抗萎缩蛋白(mM)表达载体在肌细胞去负荷时的表达水平及抗萎缩活性。方法:首先,构建了PM调控下的荧光素酶表达载体(pGL3-PM-Luc),检测模拟失重下荧光素酶基因在肌细胞中的表达水平,验证PM对重力负荷变化的响应。随后,构建了PM调控下的mM表达载体‘pGL3-PM-mM’,观察模拟失重下mM在肌细胞中的表达及其对肌细胞增殖的影响。结果:模拟失重下,‘pGL3-PM-Luc’转染的肌细胞荧光素酶表达提高了29%,‘pGL3-PM-mM’载体转染的肌细胞,mM表达上调了2.6倍,细胞增殖活性提高了24%。结论:PM在失重条件下可开启下游基因的表达,依此特点构建的mM表达载体,其mM的表达能随负荷减少而增加,且可促进肌细胞的增殖活性。Objective：To observe the ability of myostatin promoter（PM） responding to weightlessness and to explore the anti atrophy activi- ty of expression vector in which anti muscular atrophy protein ＇ mutated myostatin propeptide＇ （ mM） was constructed under control of PM. Method： PM gene was cloned, and luciferase xpression vector controled under PM was constructed （ pGL3 - PM - Lue ） to demonstrate the ability of PM responding to weightlessness. Then mM expression vector controled under PM was constructed （pGL3 - PM - raM）. pGL3 -PM -Luc and pGL3 -PM -mM were transferred into C2C12 muscle cell respectively. Result： Under simulated weightlessness condi- tion ,the expression of luciferase in C2C12 cell transfected with ＇ pGL3 - PM - Lue＇ increased 29%. The expression of mM in C2C12 cell transfected with ＇ pGL3 - PM - mM＇ elevated 2. 6 fold and the proliferation of C2C12 transfected with ＇ pGL3 - PM - raM＇ increased 24% under simulated weightlessness. Conclusion ： PM can respond to unloading signal of weightlessness. ＇ pGL3 - PM -mM ＇ vector showed the ability to express mM and subsequently to improve C2C12 proliferation under simulated weightlessness condition.

关 键 词：肌肉萎缩 肌肉生长抑制素 肌肉生长抑制素前肽 启动子 失重 

分 类 号：R852.22[医药卫生—航空、航天与航海医学]