| 富亮氨酸重复超家族新成员LRRC4的IgC2结构域蛋白的原核表达和纯化 |
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| 引用本文： | 董利,谭琛,王洁如,钱骏,张小慧,李小玲,李桂源.富亮氨酸重复超家族新成员LRRC4的IgC2结构域蛋白的原核表达和纯化[J].生命科学研究,2002,6(2):147-151. |
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| 作者姓名： | 董利 谭琛 王洁如 钱骏 张小慧 李小玲 李桂源 |
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| 作者单位： | 中南大学,湘雅医学院,肿瘤研究所,中国湖南,长沙,410078 |
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| 基金项目： | 国家自然科学基金项目 ( 30 10 0 191) |
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| 摘 要： | 富亮氨酸重复超家族新成员LRRC4基因是新克隆的脑瘤相关基因，采用多聚酶链式反应（PCR）方法获得长约500bp含IgC2结构域的DNA序列，扩增产物克隆至pGEX-4T-2质粒中，构建GST融合表达质粒，在大肠杆菌中诱导表达融合蛋白，经包涵体沉淀，溶解，Glutathione-Sepharose亲和层析纯化获得融合蛋白，并以Western blot鉴定证实，通过IgC2结构域蛋白的纯化分离该结构域，为进一步研究该结构域及LRRC4基因的结构和功能奠定了基础。
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| 关 键 词： | LRRC4 原核表达 谷胱甘肽巯基转移酶 基因表达 融合蛋白 蛋白纯化 IgC2结构域蛋白 富亮氨酸重复超家族 |
| 文章编号： | 1007-7847(2002)02-0147-05 |
| 修稿时间： | 2002年3月11日 |
Expression and Purification of IgC2 Domain in LRRC4,a Novel Member of Leucine-Rich Repeat(LRR) Superfamily |
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| DONG Li,TAN Chen,WANG Jie-ru,QIAN Jun,ZHANG Xiao-hui,LI Xiao-ling,LI Gui-yuan.Expression and Purification of IgC2 Domain in LRRC4,a Novel Member of Leucine-Rich Repeat(LRR) Superfamily[J].Life Science Research,2002,6(2):147-151. |
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| Authors: | DONG Li TAN Chen WANG Jie-ru QIAN Jun ZHANG Xiao-hui LI Xiao-ling LI Gui-yuan |
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| Abstract: | DNA sequence of IgC2 domain derived from LRRC4, a novel member of Leucine-Rich Repeat (LRR) superfamily, was obtained by PCR. A 500 bp sequence was cloned into expression vector pGEX-4T-2 to construct the GST fusion protein. The recombinant plasmid was transformed into E.coli BL21 and the over-expressed fusion protein of IgC2 domain was existed in inclusion bodies. After the inclusion bodies were precipitated and dissolved in 0.3% SKL, the soluble fusion protein was purified with glutathione affinity resin. Fusion protein was testified by Western blot using anti-GST monoclonal antibody. This work provides us the foundation to do further investigation on the structure and function of this domain. |
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| Keywords: | GST fusion protein gene expression inclusion body |
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