| 人整合素分子α4亚基片段的克隆和表达 |
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| 引用本文： | 刘永全,高杰英,罗振革,孔祥英,梅俊杰,彭虹.人整合素分子α4亚基片段的克隆和表达[J].细胞与分子免疫学杂志,2001,17(1):13-15. |
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| 作者姓名： | 刘永全 高杰英 罗振革 孔祥英 梅俊杰 彭虹 |
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| 作者单位： | 军事医学科学院微生物流行病研究所免疫室 , |
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| 基金项目： | 国家自然科学基金资助项目，No.
39789010 |
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| 摘 要： | 目的克隆并表达人整合素分子α4亚基1.2kbcDNA片段。方法从HL-60细胞系中提取总RNA，以RT-PCR法扩增人整合素分子α4亚基片段1.2kbcDNA(1753～2934bp)，并将该片段亚克隆至表达载体pGEX-3X，用IPTG诱导表达。结果克隆了α4亚基1.2kbcDNA片段。序列分析表明，其所编码的氨基酸序列与文献相比较只有一处错义突变(Arg→Gln)，经分析该突变对抗原性影响不大。将该片段亚克隆至表达载体pGEX-3X，经IPTG诱导在大肠杆菌中获得高效表达。结论获得了α4亚基1.2kbcDNA片段及其原核表达产物，对研究α4整合素的功能具有重要的意义。
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| 关 键 词： | 整合素 α4亚基 RT-PCR cDNA克隆 |
| 文章编号： | 1007-8738(2001)01-013-03 |
| 修稿时间： | 2000年1月11日 |
Cloning and expression of
the 1.2kb cDNA fragment of human integrin |
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| LIU Yong-quan,GAO Jie-ying,LUO Zhen-ge,KONG Xiang-ying,MEI Jun-jie,PENG Hong.Cloning and expression of
the 1.2kb cDNA fragment of human integrin[J].Journal of Cellular and Molecular Immunology,2001,17(1):13-15. |
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| Authors: | LIU Yong-quan GAO Jie-ying LUO Zhen-ge KONG Xiang-ying MEI Jun-jie PENG Hong |
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| Abstract: | Aim To clone and express the 1.2kb cDNA fragment (1/753-2 934bp) of human integrin α 4 subunit. Methods The 1.2 kb cDNA fragment of human integrin α 4 subunit was amplified from HL-60 total RNA by RT-PCR, then it was subcloned into expression vector pGEX-3X and induced with IPTG. Results The 1.2 kb cDNA fragment of human integrin α 4 subunit was cloned. The sequencing indicated that there was only one missense mutation (Arg→ Gln) among the fragment, and this mutation won't affect antigenicity after analysed by GOLDKEY. Then the 1.2 kb cDNA was subcloned into expression vector pGEX-3X. The α 4 fragment was highexpressed in E.coli after induced with IPTG. Conclusion The 1.2kb cDNA fragment of α 4 subunit was obtained, and it was highexpressed in E.coli, it might be important for study on the function of α 4 integrins. |
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