| 细胞黏附分子1（ICAM-1）ScFv基因原核载体的构建及表达 |
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| 引用本文： | 李月红,张国力,岳玉环,吴广谋,朱平. 细胞黏附分子1（ICAM-1）ScFv基因原核载体的构建及表达[J]. 中国兽医学报, 2012, 32(5): 725-727 |
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| 作者姓名： | 李月红 张国力 岳玉环 吴广谋 朱平 |
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| 作者单位： | 1. 吉林农业大学动物科技学院,吉林长春130118／军事医学科学院军事兽医研究所,吉林长春130122
2. 军事医学科学院军事兽医研究所,吉林长春,130122 |
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| 基金项目： | 国家自然科学基金资助项目(30972191);吉林省科技发展计划资助项目(20100232) |
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| 摘 要： | 应用PCR技术扩增ICAM-1ScFv基因,经纯化测序,得到约750bp的核苷酸片段,并构建了原核重组表达质粒pET20b-ICAM-1ScFv。将重组质粒转化至表达菌BL21（DE3）中,诱导后的SDS-PAGE分析显示,pET20b-ICAM-1ScFv表达量占菌体蛋白总量的18.4%。本试验将重组蛋白的表达条件进行优化,结果表明,ICAM-1ScFv蛋白在pET20b表达载体中的最佳表达条件为37℃诱导5h。
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| 关 键 词： | ICAM-1 ScFv 原核载体 表达 |
Construction of prokaryotic expression vector and expression of ICAM-1 ScFv gene |
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| LI Yue-hong,ZHANG Guo-li,YUE Yu-huan,WU Guang-mou,ZHU Ping. Construction of prokaryotic expression vector and expression of ICAM-1 ScFv gene[J]. Chinese Journal of Veterinary Science, 2012, 32(5): 725-727 |
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| Authors: | LI Yue-hong ZHANG Guo-li YUE Yu-huan WU Guang-mou ZHU Ping |
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| Affiliation: | 1.College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;2.The Military Veterinary Institute,Academy of Military Medicine,Changchun 130122,China) |
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| Abstract: | The(ICAM-1)ScFv gene of 750 bp were amplified by PCR.The recombinant expressing plasmid pET20a-ICAM-1 ScFv were obtained and recombinant proteins was expressed in BL21(DE3).Analysis of SDS-PAGE and thin layer scanning results showed that amount of expressed recombinant proteins was 18.4% in total lysis protein of bacteria.The expression condition was improved and the result indicates that the proper factor was 37℃,induced 5 h with IPTG. |
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| Keywords: | ICAM-1 ScFv prokaryotic vector expression |
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