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	TACO cDNA的克隆、表达及TACO的F肌动蛋白交联作用
	刘长振 ; 李湘辉 ; 隋森芳 ; LIU Chang-Zhen ; LI Xiang-Hui ; SUI Sen-Fang
	2010-06-10 ; 2010-06-10
关键词	TACO 克隆与表达 肌动蛋白纤维交联 TACO, clone and expression, F-actin cross-linking Q936 Q78
其他题名	Cloning and Expression of TACO cDNA Gene in E.coli and TACO's F-actin Cross-linking Ability
中文摘要	为进一步研究TACO(tryptophan-aspartate-containing coat protein)的性质、功能及它与肌动蛋白之间的相互关系,利用逆转录PCR技术扩增出TACO的cDNA序列,并将其克隆到pGEX2T表达质粒上.重组表达质粒pGEX2TTACO在宿主菌BL21中得到稳定表达.表达产物经GlutathioneSepharoseTM亲和纯化得到融合蛋白GSTTACO.肌动蛋白纤维(Factin)结合共沉淀实验证明,融合蛋白GSTTACO保持了天然TACO蛋白的F肌动蛋白结合活性.利用肌动蛋白纤维交联(Factincrosslinking)实验和电镜技术发现,纯化后的GSTTACO蛋白具有交联肌动蛋白纤维并将其捆聚成束的能力.实验结果表明TACO很可能起组织者的作用,通过与肌动蛋白纤维的交联,将肌动蛋白纤维捆聚成束从而形成新的肌动蛋白细胞骨架.; To further investigate the properties and biological functions of TACO and the interaction between this protein and actin, the TACO cDNA obtained by RT-PCR was inserted into pGEX-2T vector and successfully expressed in E.coli BL21. The fusion protein GST-TACO was purified to a single band(57 kD) on SDS-PAGE from lysate by glutathione-Sepharose TM. F-actin co-sedimentation assay showed that the purified GST-TACO owned the F-actin binding ability of native TACO. By F-actin cross-linking analysis and electron microscopy imaging, GST-TACO was found to cross-link F-actin into bundles. These results indicate that TACO may act as an organizer to cross-link F-actin into new actin cytoskeletons.; 国家“973”项目(No.2004CB720005)~~
语种	中文 ; 中文
内容类型	期刊论文
源URL	[http://hdl.handle.net/123456789/60661]
专题	清华大学
推荐引用方式 GB/T 7714	刘长振,李湘辉,隋森芳,等. TACO cDNA的克隆、表达及TACO的F肌动蛋白交联作用[J],2010, 2010.
APA	刘长振,李湘辉,隋森芳,LIU Chang-Zhen,LI Xiang-Hui,&SUI Sen-Fang.(2010).TACO cDNA的克隆、表达及TACO的F肌动蛋白交联作用..
MLA	刘长振,et al."TACO cDNA的克隆、表达及TACO的F肌动蛋白交联作用".(2010).

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