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绿色荧光蛋白标记的人脂联素真核表达载体的构建 被引量：1

Construction and Identification of Eukaryotic pcDNA3.1/ CT- GFP-TOPO-Adiponectin Expression Vector
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摘要 目的:构建绿色荧光蛋白(GFP)标记的人脂联素(Adiponectin)真核表达质粒。方法:用本实验室已构建好的人脂联素克隆载体pMD18-T/Adiponectin为模板,结合已设计好的特异性引物,采用PCR方法克隆出Adiponectin目的片段,将该目的片段再连至真核表达载体pcDNA3.1/CT-GFP-TOPO上,转化入TOP10感受态细胞中,通过筛选得到融合有绿色荧光蛋白的重组质粒pcDNA3.1/CT-GFP-TOPO-Adiponectin。结果:PCR获得长度为736bp的目的片段,经pcDNA3.1/CT-GFP-TOPO真核表达载体连接、筛选及序列分析后,证实所插入的目的片段与genebank检索的人脂联素cDNA序列(AccessionNM-004797)100%匹配。结论:绿色荧光蛋白标记的人脂联素真核表达载体构建成功。 Obiective： To construct the pcDNA3.1/CT -GFP-TOPO-Adiponectin expression vector. Methods： The encoding fragment of human Adiponectin gene was obtained from a recombinant plasmid pMD18-T/Adiponectin using PCR, then the gene was inserted into the vector pcDNA3.1/CT-GFP-TOPO after sequencing, which introduced into TOP10 competent ceils in order to choose and sequence, Results： A 736 bp fragment was obtained by PCR clone after bloting and sequencing. The results were matching to the cDNA of human adiponectin, which accession of the gene bank was NM-004797. Conclusion： The eukaryotic pcDNA3.1/CT-GFP-TOPO- Adipon ectin expression vector was constructed successfully.
作者 刘德敏 杜春红 孙颖 张捷
机构地区 天津医科大学代谢病医院
出处 《天津医药》 CAS 北大核心 2007年第5期324-325,I0003,共3页 Tianjin Medical Journal
基金 天津市教委基金项目(项目编号:2003025) 天津市自然科学基金资助项目(项目编号:043609211)
关键词 脂类 遗传载体 质粒 绿色荧光蛋白质类 聚合酶链反应 lipids genetic vectors plasmids green fluorescent proteins polymerase chain reaction
分类号 R512.6 [医药卫生—内科学]
  • 相关文献

参考文献5

  • 1Arita Y,Kihara S,Ouchi N,et al.Paradoxical decrease of an adipose-specific protein,adiponectin,in obesity[J].Biochem Biophys Res Commun,1999,257:79-83.
  • 2Fruebis J,Tsao T,Javorschi S,et al.Proteolycic cleavage product of 30 kDa adipocyte complement-related increases fatty acid oxidation in muscle and causes weight loss in mice[J].Proc Natl Acad Sci USA,2001,98(4):2005-2010.
  • 3刘德敏,靳立忠,孙颖,张捷.人脂联素cDNA的克隆及序列分析[J].天津医药,2005,33(2):71-73. 被引量：5
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二级参考文献13

  • 1Weyer C, Funahashi T, Tanaka S, et al. Hypoadiponectinemia in obesity and type 2 diabetes: close association with insulin resistance and hyperinsulinemia. J Clin Endocrinol M etab, 2001, 86:1930-1935.
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  • 3Takahashi M, Arita Y, Yamagata K, et al. Genomic structure and mutations in adipose-specific gene, adiponeetin, Int J Obes, 2000, 24: 861-868.
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  • 5Arita Y, Kihara S, Ouchi N, et al. Parodoxical decrease of an adipose-specific protein, adiponectin, in obesity. Biochem Biophys Res Commun, 1999, 257:79-83.
  • 6Weyer C, Funahashi T, Tanaka S, et al. Hypoadiponectinemia in obesity and type 2 diabetes: close association with insulin resistance and hypefinsulinemia. J Clin Endocfinol Metab, 2001, 86:1930-1935.
  • 7Fruebis J, Tsao TS, Javorschi S, et al. Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice.Proc Natl Acad sci USA, 2001, 98:2005-2010.
  • 8Berg AH, Combs TP, Du X, et al. The adipocyte-secreted protein Acrp30 enhances hepatic insulin action. Nat Med, 2001, 7:947-953.
  • 9Combs TP, Berg AH, Obicis, et al. Endogenous glucose prodution is inhibited by the adipose-derived protein Acrp 30. J Clin Invest,2001, 108(12): 1875-1881.
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共引文献4

同被引文献5

  • 1刘德敏,靳立忠,孙颖,张捷.人脂联素cDNA的克隆及序列分析[J].天津医药,2005,33(2):71-73. 被引量：5
  • 2Qi Y, Takahashi N, Heliman SM, et al. Adiponectin acts in the brain to decrease body weight[J]. Nat Med, 2004,10(5) : 524-529.
  • 3Lindmark S,Buren J,Eriksson JW. Insulin resistance,endocrine function and adipokines in type2 diabetes patients at different glycaemic levels: potential impact for glucotoxicity in vivo [J]. Clin Endocrinol (Oxf), 2006,65(3) :301-309.
  • 4Manigrasso MR, Ferroni P, Santilli F,et al. Association between circulating adiponectin and interleukin-10 levels in android obesity: effects of weight loss [J]. Clin Endocrinol Metab,2005,90 (10): 5876-5879.
  • 5Kadowaki T,Yamauchi T,Kubota N,et al. Adiponectin and adipon ectin receptors in insulin resistance, diabetes, and the metabolic syndrome[J]. J Clin Invest, 2006,116( 7 ) : 1784-1792.

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二级引证文献1

天津医药
2007年 第5期

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