肌球蛋白X干涉载体的构建与鉴定  

Construction and Identification of Myosin X Interference Vector

作　　者：王俊杰[1,2] 丁伟[1] 于龙[1] 朱磊[1] 

机构地区：[1]曲靖师范学院生物资源与环境科学学院,曲靖655011 [2]东北师范大学细胞与遗传研究所,长春130024

出　　处：《生物技术通报》2011年第2期192-196,共5页Biotechnology Bulletin

基　　金：云南省教育厅科学研究基金项目(2010Y080);曲靖师范学院青年基金项目(2009QN020)

摘　　要：肌球蛋白X(Myosin X,Myo X)是近年来发现的在细胞运动中发挥重要功能的非传统型肌球蛋白,但其发挥功能的分子机制目前尚不明确,RNA干扰技术是目前研究基因功能常用的方法。构建针对鼠的Myo X基因mRNA的shRNA真核表达载体,并转染NLT细胞,观察其对Myo X表达的抑制效应。结果显示,shRNA-2能够有效地降低Myo X的蛋白表达水平,而shRNA-1则不能。进一步分析显示,转染shRNA-2的NLT细胞丝状伪足的形成受到抑制,表明shRNA-2是能够抑制Myo X表达的干涉载体,此干涉载体的构建为进一步研究Myo X的新功能,揭示其作用的分子机制奠定基础。Myosin X is an unconventional myosin which have a role in the cellular motility,but the associated molecular mechanism is not well known.RNAi（RNA interference） is a powerful tool presently known in the study of gene function.In this study,a recombinant vector carry the shRNA of Myo X was constructed and transferred to NLT cells.The expression of Myo X was suppressed by shRNA-2 but shRNA-1 did not have obvious effect on the expression level.Immunofluorescence assay showed that the filopodial formation was suppressed in NLT cells transfected with shRNA-2.Therefore,vector shRNA-2 is efficient in the interfering Myo X expression which would be useful in the study of Myo X.

关 键 词：肌球蛋白X RNA干涉 丝状伪足 SHRNA 表达水平 

分 类 号：Q78[生物学—分子生物学]