摘要
The ribosomal protein S29 also known as RPS29, is not only a component of the 40S subunit of ribosome, but also involved in embryonic development, oncogenesis and other pathologic conditions. However, rare commercial antibody against RPS29 restricts the discovery of precise physiological and pathological function of this protein. In this study, the whole RPS29 gene was inserted into plasmid pGEX-6p-1 to express glutathione＇s transferase （GST） fusion proteins in Escherichia eoli （E. coli） strain BL21. High yields of soluble recombinant proteins were obtained. Mice were immunized with the recombinant RPS29 protein. The serum from the immunized mice could specially react with purified recombinant RPS29 proteins and native RPS29 proteins in CCE cells by western blotting, immunofluorescence staining and flow cytometric analysis. Further more the polyclonal antibodies also reacted specifically with human cell strain ECV304, which showed typical cytoplasmatic fluorescence. The polyclonal antibodies we prepared would be an available tool for studying the roles of RPS29 in embryonic development and human diseases.
也作为 RPS29 知道的 ribosomal 蛋白质 S29，是核糖体的 40S 子单元的一个部件不仅，而且在胚胎的开发， oncogenesis 和另外的病理学的条件包含了。然而，对 RPS29 的稀罕商业抗体限制这蛋白质的精确生理、病理学的功能的发现。在这研究，整个 RPS29 基因被插入到 plasmid pGEX-6p-1 表示谷胱甘肽的 transferase (GST ) 在 Escherichia coli 的熔化蛋白质(E。coli ) 紧张 BL21。可溶的 recombinant 蛋白质的高收益被获得。老鼠与 recombinant RPS29 蛋白质被使免疫。从使免疫的老鼠的浆液能特殊由西方的弄污， immunofluorescence 染色和流动 cytometric 分析在 CCE 房间与净化的 recombinant RPS29 蛋白质和本国的 RPS29 蛋白质反应。进一步更 polyclonal 抗体也与人的房间紧张 ECV304 明确地反应了，它显示出典型 cytoplasmatic 荧光。我们准备了的 polyclonal 抗体将是为在胚胎的开发和人的疾病学习 RPS29 的角色的一个可得到的工具。
基金
Supported by the National Natural Science Foundation of China(30800983,30700418 and 30972596)
the Natural Science Foundation of Chongqing(2008BB5113 and 2009BB5015)
the Scientific Research Foundation of Third Military Medical University(2009XHG03 and 2009XYY04)
