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人胰高血糖素样肽-1突变体基因原核表达质粒的构建

Construction of expression vector of human Glucagon-like peptide-1 mutant gene
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摘要 目的:对人胰高血糖素样肽-1(hGLP-1)进行基因突变,构建GST融合蛋白原核表达质粒pGEX-4T/(Val8)hGLP-1,并在大肠杆菌中诱导表达。方法:选择大肠杆菌偏爱密码子,将hGLP-1(7～37)第2位的丙氨酸(Ala8)定点突变为缬氨酸(Val8),直接生物合成4个寡聚核苷酸片段并进行基因拼接,形成完整的hGLP-1突变体基因(Val8)hGLP-1,并连接到载体pGEX-4T-1,转化大肠杆菌BL21,BamH I与Xho I双酶切电泳鉴定与序列测定,然后诱导表达其融合蛋白。结果:经酶切电泳鉴定与测序分析,证实所合成的突变体基因(Val8)hGLP-1已克隆到pGEX-4T-1中,经SDS-PAGE分析可以诱导表达出融合蛋白。结论:成功地构建了GST融合蛋白原核表达质粒pGEX-4T/(Val8)hGLP-1,为进一步获得重组hGLP-1蛋白奠定基础,为产业化规模制备hGLP-1突变体提供实验依据。 Objective: To obtain the mutant gene of human Glucagon-like peptide-1(hGLP-1),construct the expression vector of pGEX-4T/(Val8)hGLP-1,and express the GST fusion protein in E.coli cells.Methods: With the preferential codon of E.coli being selected,four oligonucleotide fragments were directly biosynthesized,then were spliced to form a complete mutant gene of hGLP-1 which changed the codon for the second amino acid of hGLP-1(7~37)-alanine(Ala8) into valine(Val8) by means of site-directed mutagenesis,so it name...
作者 朱清 顾云 陆伟 刘梅
机构地区 南通大学医学院药学教研室 江苏省神经再生重点实验室
出处 《南通大学学报（医学版）》 2008年第3期165-168,共4页 Journal of Nantong University(Medical sciences)
基金 南通大学自然科学研究资助项目(05Z084)
关键词 人胰高血糖素样肽-1 融合蛋白 基因突变 克隆 糖尿病 Human Glucagon-like peptide-1 Fusion protein Gene mutation Cloning Diabetes mellitus
分类号 R587.1 [医药卫生—内分泌]
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参考文献10

  • 1[1]Vilsboll T,Krarup T,Madsbad S,et al.No reactive hypo-glycaemia in type 2 diabetic patients after subcutaneous administration of GLP-1 and intravenous glucose[J].Diabet Med,2001,18(2):144-149.
  • 2乐嘉静,李湛君,邱财荣,宫泽辉,徐康森.重组人胰高血糖素类多肽-1(7-36)对正常动物降糖作用的实验研究[J].中国临床药理学与治疗学,2004,9(5):527-531. 被引量：7
  • 3[3]De L,Diva D,Deng SP,et al.Role of endogenous glucagons-like peptide-1 in islet regeneration after partial pancreatectomy[J].Diabetes,2003,52(2):365-371.
  • 4[4]Zander M,Medsbad S,Madsen JL,et al.Effect of 6-week course of glucagons-like peptide-1 on glycaemic control,insulin sensitivity,and β-cell function in type 2 diabetes:as parallel -group study[J].Lancet,2002,359(9309):824-830.
  • 5[5]Neumiller JJ,Odegard PS,White JR Jr,et al,looking to the future:focus on DPP-4 inhibitors for the treatment of type 2 diabetes and emerging therapies[J].Diabetes Edu-cation,2008,34(2):183-200.
  • 6张志珍,毛积芳,窦鸿,杨生生.人胰高血糖素样肽-1 cDNA的克隆与表达[J].第二军医大学学报,2001,22(4):316-318. 被引量：7
  • 7[7]Green BD,Gault VA,O'harte FP,et al.Structurally modi-fied analogues of glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) as fu-ture antidiabetic agents[J].Curr Pharm Des,2004,10(29):3651-3662.
  • 8[8]Gallwitz B,Ropeter T,Morys-Wortmann C,et al.GLP-1-analogues resistant to degradation by dipeptidyl-peptidase Ⅳ in vitro[J].Regul Pept,2000,86(1-3):103-111.
  • 9[9]Green BD,Mooney MH,Gault VA,et al.N-terminal His(7)-modification of glucagon-like peptide-1 (7-36) amide generates dipeptidyl peptidase Ⅳ-stable analogues with potent antihyperglycaemie activity[J].Journal of En-docrinology,2004,180(3):379-388.
  • 10尹晓敏,施建华,刘飞.剪接因子SRp55原核表达质粒的构建和表达[J].南通大学学报（医学版）,2007,27(2):79-82. 被引量：1

二级参考文献20

  • 1萨姆布鲁克J 弗里奇EF.分子克隆实验指南（第2版）[M].北京:科学出版社,1995.565-571.
  • 2[1]Brubaker PL. Regulation of intestinal proglucagon-derived peptide secretion by intestinal regulatory peptides[J]. Endocrinology,1991;128(2):3175-28
  • 3[2]Reberge JN, Brubaker PL. Regulation of intestinal proglucagon-derived peptide secretion by glucose-dependent insulinotropic peptide in a novel enteroendocrine loop [J]. Endocrinology,1993;133(1):233-40
  • 4[3]Orskov C, Holst JJ, Knuhtesen S. Glucagon-like peptide GLP-1 and GLP-2,predicted products of the glucagons gene,aresecreted separately from pig small intestine but not pancreas[J].Endocrinology,1986;119(1):1467
  • 5[4]Fehmann HC, Habener JF. Insulinotropic hormone glucagons-like peptide-1(7-37)stimulation of proinsulin gene expression and proinsulin biosynthesis in insulinom a β TC-1 cells[J].Endocrinol,1992;130(3):159-66
  • 6[5]Mojsov S, Weir GC, Habener JF. Insulinotropin: glucagons-like peptide 1 (7-37) co encoded in the glucagons gene is apotent stimulator of insulin release in the perfused rat panicreas[J]. J Clin Invest,1987;79(2):616
  • 7[1]Maglott D,Ostell J,Pruitt KD,et al.Entrez Gene:genecentered information at NCBI[J].Nucleic Acids Research,2005,33 (Database issue):D54-8.
  • 8[2]Pruitt KD,Tatusova T,Maglott DR.NCBI reference sequence:a curated non-redundant sequence database of genomes,transcripts and proteins[J].Nucleic Acids Research,2005,33 (Database issue):D501-504.
  • 9[3]Graveley BR.Sorting out the complexity of SR protein functions[J].RNA,2000,6:1197-1201.
  • 10[4]Manley JL Tacke R.SR proteins and splicing control[J].Genes Dev,1996,10:1569-1572.

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南通大学学报（医学版）
2008年 第3期

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