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| Jo-1蛋白在大肠杆菌中的克隆与表达 | |
| 引用本文： | 辛海涛,赵晓瑜,刘国振,王卫宁,侯颖娜,张峰. Jo-1蛋白在大肠杆菌中的克隆与表达[J]. 河北农业大学学报, 2006, 29(5): 69-71,75 |
| 作者姓名： | 辛海涛 赵晓瑜 刘国振 王卫宁 侯颖娜 张峰 |
| 作者单位： | 河北大学,生命科学学院,河北,保定,071002;中科院基因组研究所,北京,100009;保定市建设医院,河北,保定,071000 |
| 摘 要： | 构建在E.coli中表达人组氨酰-tRNA合成酶(Jo-1)的质粒。用RT-PCR从人胎盘组织总RNA中获得编码人组氨酰-tRNA合成酶(Jo-1)基因的全序列,将此片段定向克隆到麦芽糖融合蛋白表达载体pMAL-c中,诱导表达获得Jo-1的可溶性融合蛋白,并通过亲和层析予以纯化。结果免疫印迹实验表明,表达的融合蛋白能与含有抗Jo-1抗体的病人血清发生特异反应,重组融合蛋白具有Jo-1蛋白的免疫原性和免疫特异性。 |
| 关 键 词： | 人组氨酰-tRNA合成酶 PCR 克隆 表达 自身抗原 |
| 文章编号： | 1000-1573（2006）05-0069-03 |
| 收稿时间： | 2006-03-23 |
| 修稿时间： | 2006-03-23 |
Cloning and expression of Jo1 protein in E.coli |
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| XIN Hai-tao,ZHAO Xiao-yu,LIU Guo-zhen,WANG Weining,HOU Ying-na,ZHANG Feng. Cloning and expression of Jo1 protein in E.coli[J]. Journal of Agricultural University of Hebei, 2006, 29(5): 69-71,75 | |
| Authors: | XIN Hai-tao ZHAO Xiao-yu LIU Guo-zhen WANG Weining HOU Ying-na ZHANG Feng |
| Affiliation: | 1. College of Life Science, Hebei University, Baoding 071002, China; 2. Beijing Genomics Institute, Beijing 100009, China; 3.Construction Hospital of Baoding, Baoding 071000, China |
| Abstract: | A vector expressing human histidyl-tRNA synthetase(Jo-1) protein in E.coli was constructed.The full-length DNA fragment encoding Jo-1 protein was obtained by RT-PCR from human placenta tissue.The Jo-1 fragment was cloned into plasmid pMAL-c(a maltose-binding protein,MBP fusion vector) and expressed in E.coli.Western-blotting showed that the fusion protein could react with anti-MBP antibody and anti-Jo-1 antibodies of patients serum,but not with normal serum.Jo-1 protein has immune specificity. |
| Keywords: | Jo-1 protein PCR cloning expression self antigen |
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