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肌球蛋白X干涉载体的构建与鉴定
引用本文:王俊杰,丁伟,于龙,朱磊.肌球蛋白X干涉载体的构建与鉴定[J].生物技术通报,2011(2).
作者姓名:王俊杰  丁伟  于龙  朱磊
作者单位:1. 曲靖师范学院生物资源与环境科学学院,曲靖,655011;东北师范大学细胞与遗传研究所,长春,130024
2. 曲靖师范学院生物资源与环境科学学院,曲靖,655011
基金项目: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。

关 键 词:肌球蛋白X  RNA干涉  丝状伪足  shRNA  表达水平  

Construction and Identification of Myosin X Interference Vector
Wang Junjie,Ding Wei,Yu Long,Zhu Lei.Construction and Identification of Myosin X Interference Vector[J].Biotechnology Bulletin,2011(2).
Authors:Wang Junjie  Ding Wei  Yu Long  Zhu Lei
Institution:Wang Junjie1,2 Ding Wei1 Yu Long1 Zhu Lei1(1College of Biology and Enviromental Sciences,Qujing Normal University,Qujing 655011,2Institute of Cytology and Genetics,Northeast Normal University,Changchun 130024)
Abstract: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 su...
Keywords:Myosin X RNA interference Filopodia shRNA Expression level  
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