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人脂联素基因的克隆及在大肠杆菌中的表达 被引量：1

Study on the Clone of Human Adiponectin Gene and Its Expression in Escherichia Coli
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摘要 目的:克隆人脂联素(adiponectin)基因,并在大肠杆菌宿主系统中高效表达出adiponectin蛋白进行鉴定。方法:从人皮下脂肪组织提取总RNA,确认其正确性后经RT-PCR扩增得到全长cDNA片段,此片段经纯化回收后克隆至pMD18-T载体,经行DNA序列分析,进一步克隆构建到表达载体至pET-DEST42中,用IPTG在大肠杆菌中诱导表达。结果:含重组adiponectin质粒的大肠杆菌经0.4mmol/L的IPTG诱导6h后有高表达。结论:adiponectin基因的克隆构建和在大肠杆菌中的表达获得成功。 Objective： To clone human adiponectin gene and identify its expression in E.coli. Methods： Total RNA was extracted from human subcutaneous adipose tissue and a full length cDNA was obtained by RT-PCR. The product was ligated to pMD-18T vector, and was sequenced and constructed to the expression vector in pET-DEST 42, and the expression was induced by IPTG in E.coli. Results： A high level expression was obtained by 04mmol/L IPTG inducement in recombined adiponectin E. coil Conclusion： Human adiponectin gene in E.coli is successfully cloned and expressed.
作者 刘德敏 杨莉丽 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 2006年第3期148-150,共3页 Tianjin Medical Journal
基金 天津市自然科学基金资助项目(项目编号:20030325)
关键词 胞间信号肽类和蛋白质类 克隆 分子 基因表达 大肠杆菌 intercellular signaling peptides and proteins cloning, molecular gene expression escherichia coli
分类号 R589.2 [医药卫生—内分泌] R378.21 [医药卫生—病原生物学]
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参考文献3

  • 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
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二级参考文献13

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共引文献4

同被引文献11

  • 1杨泽华,解军,杨琦,张悦红,牛勃.人脂联素球状结构域(gAd)基因的克隆、融合表达和纯化[J].中国生物制品学杂志,2006,19(1):17-20. 被引量：2
  • 2Miczke A, Bryl W, Szuliaska M, et al. The role of adiponectin in pathogenesis of insulin resistance and metabolic syndrome[J]. Pol Merkuriusz Lek, 2005, 19 (113): 723.
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  • 6封淑颖 邹竹荣.人脂联素基因的克隆及其在大肠杆菌中的高效表达.中国生物工程杂志,2006,26:15-15.
  • 7LaVallie E R, Lu Z, Diblasio-Smith E A, etal. Thioredoxin as a fusion partner for production of soluble recombinant proteins in Escherichia coli [J]. Methods Enzymol, 2000, 326: 322.
  • 8Turner P, Holst O, Karlsson E N. Optimized expression of soluble cyclomaltodextrinase of thermophilic origin in Escherichia coli by using a soluble fusion-tag and by tuning of inducer concentration [J]. Protein Expr Purif, 2005, 39(1) : 54.
  • 9Malakhov M P, Mattern M R, Malakhova O A, et al. SUMO fusions and SUMO-speeifie protease for efficient expression and purification of proteins [J]. J Struet Funct Genomies, 2004, 5 (1,2): 75.
  • 10Kohli B M, Ostermeier C. A Rubredoxin based system for screening of protein expression conditions and on-line monitoring of the purification process [J]. Protein Expr Purif, 2003, 28 (2) : 362.

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天津医药
2006年 第3期

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