| 晚期糖基化终末产物受体真核表达载体的构建与表达 |
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| 引用本文： | 李煜生,龚小卫,程蔚蔚,魏洁,姜勇. 晚期糖基化终末产物受体真核表达载体的构建与表达[J]. 南方医科大学学报, 2007, 27(7): 983-986 |
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| 作者姓名： | 李煜生 龚小卫 程蔚蔚 魏洁 姜勇 |
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| 作者单位： | 南方医科大学病理生理学教研室和广东省蛋白质组学重点实验室,广东,广州,510515;南方医科大学病理生理学教研室和广东省蛋白质组学重点实验室,广东,广州,510515;南方医科大学病理生理学教研室和广东省蛋白质组学重点实验室,广东,广州,510515;南方医科大学病理生理学教研室和广东省蛋白质组学重点实验室,广东,广州,510515;南方医科大学病理生理学教研室和广东省蛋白质组学重点实验室,广东,广州,510515 |
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| 基金项目： | 国家高技术研究发展计划(863计划) , 广东省科技厅科技计划 , 广东省自然科学基金 , 广东省广州市科技计划 |
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| 摘 要： | 目的 构建带HA标签的晚期糖基化终末产物受体真核细胞表达载体.方法 采用PCR方法从人cDNA文库中扩增RAGE基因编码区,使用常规酶切、连接方法将其重组至pcDNA3真核表达载体中,对阳性克隆进行酶切、PCR和测序鉴定.以RAGE/pcDNA3重组质粒为模板,运用突变方法在RAGE氨基末端信号肽后插入HA序列.利用Western blotting在HEK 293细胞中对经过测序鉴定的HA-RAGEpcDNA3重组质粒的表达进行了检测.结果 RAGE/pcDNA3重组质粒经酶切、PCR和测序鉴定正确无误,HA-RAGEpcDNA3重组质粒可在HEK 293细胞中表达.结论 成功构建了带HA标签的RAGE真核细胞表达载体,该载体能在真核细胞中表达,为进一步研究RAGE在细胞信号转导途径中的作用提供了一个重要的工具.
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| 关 键 词： | RAGE 载体构建 基因表达 |
| 文章编号： | 1673-4254（2007）07-0983-04 |
| 修稿时间： | 2007-02-04 |
Construction and expression of the fusion vector for HA-tagged human RAGE gene |
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| LI Yu-sheng,GONG Xiao-wei,CHENG Wei-wei,WEI Jie,JIANG Yong. Construction and expression of the fusion vector for HA-tagged human RAGE gene[J]. Journal of Southern Medical University, 2007, 27(7): 983-986 |
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| Authors: | LI Yu-sheng GONG Xiao-wei CHENG Wei-wei WEI Jie JIANG Yong |
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| Affiliation: | Department of Pathophysiology and Key Laboratory of Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515, China |
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| Abstract: | Objective To construct a mammalian expression vector for HA-tagged receptor of advanced glycation end products(RAGE).Methods Human RAGE cDNA codon region was amplified by PCR from human cDNA library and cloned into the pcDNA3 vector following routine procedures.After identification by enzyme digestion,PCR and sequencing,the correct recombinant vector RAGE/pcDNA3 was inserted with HA sequence behind the signal peptide sequence of RAGE.After identification by sequencing,HA-RAGEpcDNA3 was transfected into HEK293 cells,and its expression was detected by Western blotting.Results Identification by enzyme digestion,PCR and sequencing,confirmed the validity of the recombinant vector RAGE/pcDNA3,and HA-RAGEpcDNA3 was highly expressed in HEK 293 cells.Conclusion HA-tagged RAGE is successfully constructed and expressed in mammalian cells,which may facilitate functional study of RAGE in cell signal transduction. |
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| Keywords: | receptor, advanced glycation end products vector construction gene expression |
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