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重组真核表达载体pEGFP-BDNF的构建 被引量：1

Construction of pEGFP-BDNF
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摘要 目的将脑源性神经生长因子(BDNF)构建到表达质粒(pEGFP-N1)上。方法本实验采用RT-PCR的方法,从大鼠海马总RNA中扩增目的基因,分别用一步法(直接构建到表达载体上)和两步法(先构建到克隆载体再构建到表达载体上)构建重组载体,通过酶切和基因测序筛选鉴定正确的阳性克隆,并对两种方法做简单的比较。结果两种方法构建的重组载体都插入了正确的序列。两步法的转染效率明显高于一步法,但一步法也同样得到了正确的阳性克隆。结论成功构建了pEGFP-BDNF重组表达载体。 Objective To construct pEGFP-BDNF.Methods BDNF gene was amplified by RT-PCR method from total RNA of rat hippocampus tissue,then the recombination vector system was constructed by one-step（constructed to expression vector directly） and two-step（construct to cloning vector firstly then to expression vector） approaches respectively.The right recombination was selected by enzyme digestion and identified by gene sequencing.Results The correct sequence was identified in both approaches.Transfection efficiency by two-step approach was much higher than one-step,but correct positive clones were also obtained by one-step.Conclusion Recombinant expression vector was constructed successfully.
作者 刘菲菲 龙大宏 龙静怡
机构地区 广州医学院人体解剖学教研室 南方医科大学
出处 《解剖学研究》 CAS 2012年第2期107-110,共4页 Anatomy Research
基金 广东省自然科学基金项目(9151063101000016) 广州市科技局应用基础研究计划项目(2009J1-C361-2)
关键词 脑源性神经生长因子 基因克隆 增强型绿色荧光蛋白 Brain-derived neurotrophic factor Gene clone Enhanced green fluorescent protein
分类号 R346 [医药卫生—基础医学]
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参考文献11

  • 1李雪琴,魏瑞丽,马晓霞.脑源性神经营养因子的研究现状及应用前景[J].中国实用神经疾病杂志,2006,9(2):45-47. 被引量：15
  • 2Blurton-Jones M, Kitazawa M, Martinez-Coria H, et al. Neural stem cells improve cognition via BDNF in a trans- genic model of Alzheimer disease. Proc Natl Acad Sci U S A, 2009,106 : 13594-13599.
  • 3Gash DM, Chen Y, Gerhardt G. Neurotrophic Factors and Parkinson's Disease. Hendb Clin Neurol, 2007,83:521- 533.
  • 4Wang Z, Deng Q, Zhang X, et al. Treatment of injured neurons with bone marrow stem ceils cotransfected by hTERT and Ad-BDNF in vitro. J Mol Neurosci, 2009, 38 : 265-272.
  • 5Fabrice G, Italina C, Charles A, et al. A transgenic mouse model engineered to investigate human brain-derived neu- rotrophic factor in vivo Transgenic Res, 2007,16 : 223-237.
  • 6Takeshi F, Seizaburo S, Michiko K. Construction of novel single-cell screening system using a yeast cell chip for nano- sized modified-protein-displaying libraries. NanoBiote- chnology, 2005,1 : 105-112.
  • 7McTigue DM, Homer PJ, Stokes BT, et al. Neurotrophin-3 and brain-derived neurotrophic factor induce oligodendro- cyte proliferation and myelination of regenerating axons in the contused adult rat spinal cord. J Neurosci, 1998,18: 5354-5365.
  • 8Liu Y, Himes BT, Solowska J, et al. Intraspinal delivery of neurotrophin-3 using neural stem ceils genetically modified by recombinant retrovirus. Exp Neurol, 1999,158 : 9-26.
  • 9Lu L, Huo J, Liu J, et al. Effect of recombinant pEGFP-N3- APC vectors carrying various APC functional domains on the expression of beta-catenin in HT-29 cells. Zhong Nan Da Xue Xue Bao Yi Xue Ban, 2010,35 : 140-145.
  • 10Li J, Zhou J, Zhong M. Construction and expression of the eukaryotic green fluorescent protein expression vector pEGFP-N1-ZNF217. Nan Fang Yi Ke Da Xue Xue Bao, 2010,30: 391-393.

二级参考文献23

  • 1[1]Kolbeck R,Bartke I,Eberle W,et al.Brain～derived neurotrophic factor levels in the nervous system of wild-type and neurotrophin rene mutant mice[J].J Neurochem,1999,72(5):1930～ 1938
  • 2[2]Butte MJ.Neurotrophic factor structures reveal clues to evolution,binding,specificity,and receptor activation[J].Cell Mol Life Sci,2001,58(8):1003～1013
  • 3[3]Ozcelik T,Rosenthal A,Foancke U,et al.Chromosomal mapping of Brain derived neurotrophic factor and neurotrophin-3 gene in man and mouse[J].Genomics,1991,10(3):569
  • 4[4]Ojeda SR,Romero C,Tapia V,et al.Neurotrophic and cell-cell dependent control of early follicular development[J].Mol Cell Endocrinol 2000,163(1～2):67～71
  • 5[5]Butowt R,Von Bartheld CS.Anterograde axonal transport of BDNF and NT-3 by retinal ganglion cells:rolls of neurotrophin receptors[J].Mol Cell Neurosci,2005,29(1):11～25.
  • 6[6]Sugimoto T,Ikuroda H,Horii Y,et al.Signal transduction pathways through TRK-A and TRK-B receptors in human neuroblastoma cells[J].J Cancer Res,2001,92(2):152～160
  • 7[7]Almeida RD,Manads BJ.Neuroprotection by BDNF against glutamate-induced apoptotic cell death is mediated by ERK and PI3～kinase pathways[J].Cell Death Diffa,2005,20
  • 8[8]Ji Y,Pang PT.Cyclic AMP controls BDNF-induced TrkB phosphorylation and dentritic spine formation in mature hippocampal neurons[J].Nat Neurosci,2005,8(2):164～172
  • 9[9]Lindsay RM,Altra CA,Cedrabaum JM,et al.The therapeutic potential of reurotrophic factors in the treatment of Parkinson' s disease[J].Exp Neurol,1993,124(I):103
  • 10[10]Furukawa K.Secreted amyloid precursor proteinαselectively suppress N-methyl-D-aspartate currents in hippocampal neurons:involvement of cyclic GMP[J].Neuroscience,1998,83:429～438

共引文献14

同被引文献14

  • 1Ni XQ,Li X,Zhang BH,et al.NGF/TrkA-mediated Kidins220/ARMS Signaling activated in the allergic airway challenge in mice.Annals of Allergy,Asthma&Immunology,2010,105:299-306.
  • 2Liu YL,Zhang BH,Fang XB,et al.NGF mediated sh2bbeta/akt signal pathway activated in allergic airway challenge in mice.Respirology,2010,15(1):80-87.
  • 3Lommatzsch M,ScMoetcke K,Klotz J,et al.Brain-deriveded neurotrophic factor in platelets and airflow limitation in asthma.Am J Respir Crit Care Med,2005,171(2):115-120.
  • 4Lommatzsch M,Lindner Y,Edner A,et al.Adverse effects of salmeterol in asthma:a neuronal perspective.Thorax,2009,64:763-769.
  • 5Prakash YS,Michael A,Christina M,et al.Brain-De-rived Neurotrophic Factor in TNF-αModulation of Ca2+in Human Airway Smooth Muscle.Am J Respir Cell Mol Bio,2009(41):603-611.
  • 6Hahn C,Islamian AP,Renz H,et al.Airway epithelial cells produce neurotrophins and promote the survival of eosinophi is during allergic airway inflammation.J Allergy Clin Immunol,2006,117(4):787-794.
  • 7Tapia Arancibia L,Rage F,Givalois L,et al.Physiology of BDNF:focus on hypothalamic function.Front Neuroendocrinol,2004,25(2):77-107.
  • 8Schramm A,Schulte JH,Astrahantseff K,et al.Biological effects of TrkA and TrkB receptor signaling in neuroblastoma.Cancer Lett,2005,228(1-2):143-153.
  • 9Ramadan BS,Richard JM,Musa AH,et al.Role of brain-derived neurotrophic factor in hyperoxia-induced enhancement of contractility and impairment of relaxation in lung parenchyma.Am J Physiol Lung Cell Mol Physiol,2008,295(2):348-355.
  • 10Kwak YG,Song CH,Yi HK,et al.Involvement of PTEN in airway hyperresponsiveness and inflammation in bronchial asthma.J Clin Invest,2003,111:1083-1092.

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解剖学研究
2012年 第2期

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