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| 融合蛋白mTSLPR-Ig的表达、纯化及其鉴定 | |
| 引用本文： | 张方,施毅,黄钢,李子玲,胡波,宋勇.融合蛋白mTSLPR-Ig的表达、纯化及其鉴定[J].细胞与分子免疫学杂志,2011,27(5):511-514. |
| 作者姓名： | 张方 施毅 黄钢 李子玲 胡波 宋勇 |
| 作者单位： | 1. 南京军区南京总医院呼吸内科,江苏,南京,210002 2. 第三军医大学分子遗传教研室,重庆,400038 3. 南京军区联勤部卫生部,江苏,南京,210002 |
| 基金项目： | 中国博士后科学基金项目，江苏省博士后科研资助计划项目 |
| 摘 要： | 目的:构建含小鼠TSLPR胞外段与小鼠Ig Fc段融合基因的腺病毒表达载体(Ad-mTSLPR-Ig),体外表达、纯化并鉴定可溶性融合蛋白mTSLPR-Ig。方法:运用PCR方法从小鼠胸腺cDNA文库中扩增mTSLPR胞外段基因,构建并鉴定mTSLPR胞外段与小鼠Ig Fc段融合基因的腺病毒表达载体(Ad-mTSLPR-Ig);将Ad-TSLPR-mIg转染COS-7细胞,收集细胞上清液;采用双抗体ELISA夹心法检测上清中融合蛋白表达;经蛋白A亲和层析,制备纯化可溶性融合蛋白mTSL-PR-Ig。结果:经测序证实,所构建表达载体的目的基因片段mTSLPR-Ig序列正确,成功包装出腺病毒表达载体Ad-mTSL-PR-Ig;ELISA法检测到上清中融合蛋白mTSLPR-Ig的表达;通过亲和层析法获得纯化的融合蛋白,经Western blot鉴定融合蛋白表达无误。结论:成功获得可溶性融合蛋白mTSLPR-Ig,为进一步研究其生物学特性奠定基础。 |
| 关 键 词： | TSLPR 可溶性受体 TSLP 哮喘 |
Expression,purification and identification of fusion protein mTSLPR-Ig |
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| ZHANG Fang,SHI Yi,HUANG Gang,LI Zi-ling,HU Bo,SONG Yong.Expression,purification and identification of fusion protein mTSLPR-Ig[J].Journal of Cellular and Molecular Immunology,2011,27(5):511-514. | |
| Authors: | ZHANG Fang SHI Yi HUANG Gang LI Zi-ling HU Bo SONG Yong |
| Institution: | ZHANG Fang1,SHI Yi1,HUANG Gang2,LI Zi-ling1,HU Bo1,SONG Yong1 1Nanjing General Hospital of Nanjing Military Region,Department of Respiratory Medicine,Nanjing 210002,2Department of Molecular Genetics,Third Military Medical University,Chongqing 400038,3Joint Logistics Department of the Nanjing Military Region,China |
| Abstract: | AIM: To construct an adenovirus vector(Ad-mTSLPR-Ig) expressing the fusion protein of the extracellular domain of mouse TSLPR and the Fc fragment of mouse Ig,further purify and assess the soluble fusion protein mTSLPR-Ig.METHODS: The gene of mTSLPR extracellular domain was amplified from cDNA library of mouse thymus by PCR.Then an adenovirus vector(Ad-mTSLPR-Ig) expressing the fusion protein of mTSLPR and the Fc fragment of mouse Ig was constructed.After Ad-mTSLPR-Ig was transfected into COS-7 cells,the sup... |
| Keywords: | TSLPR TSLP |
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