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| 携带人alpha-突触核蛋白真核重组质粒构建及其SK-N-SH细胞表达 | |
| 引用本文： | 和青,杜婷婷,宋宁,谢俊霞,姜宏. 携带人alpha-突触核蛋白真核重组质粒构建及其SK-N-SH细胞表达[J]. 青岛大学医学院学报, 2010, 46(5): 377-379,383. DOI: 10.3969/j.issn.1672-4488.2010.05.001 |
| 作者姓名： | 和青 杜婷婷 宋宁 谢俊霞 姜宏 |
| 作者单位： | 青岛大学国家生理学重点(培育)学科,山东省神经相关疾病的机制与防治重点实验室,山东,青岛,266071 |
| 基金项目： | 国家自然科学基金，山东省自然科学杰出青年基金 |
| 摘 要： | 目的构建携带人alpha-突触核蛋白真核重组质粒,检测其在人神经母细胞瘤SK-N-SH细胞中的表达及对细胞存活率的影响。方法 PCR扩增alpha-突触核蛋白基因,产物纯化后与pcDNA3.1(+)载体进行连接反应,构建pcDNA3.1(+)/alpha-突触核蛋白真核重组质粒。Lipofectamine法转染人神经母细胞瘤SK-N-SH细胞,RT-PCR方法检测SK-N-SH细胞中alpha-突触核蛋白mRNA的表达,四甲基偶氮唑盐(MTT)法检测其对细胞存活率的影响。结果酶切鉴定及基因测序证明人alpha-突触核蛋白基因已被完整、正确地插入到pcDNA3.1(+)载体中。RT-PCR结果显示该表达载体转染SK-N-SH细胞后,细胞内alpha-突触核蛋白mRNA的表达水平明显增高,MTT方法检测结果显示该表达载体转染并未引起细胞存活率的改变。结论 alpha-突触核蛋白真核重组质粒构建成功,并可在SK-N-SH细胞内表达,且对细胞的存活率不产生任何影响,从而为进一步研究alpha-突触核蛋白在帕金森病发病机制中的作用奠定了基础。 |
| 关 键 词： | α突触核蛋白 质粒 帕金森病 |
CONSTRUCTION OF THE EUKARYOTIC EXPRESSION PLASMID OF HUMAN alpha-SYNUCLEIN GENE AND ITS EXPRESSION IN SK-N-SH CELLS |
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| HE QING,DU TING-TING,SONG NING,et al. CONSTRUCTION OF THE EUKARYOTIC EXPRESSION PLASMID OF HUMAN alpha-SYNUCLEIN GENE AND ITS EXPRESSION IN SK-N-SH CELLS[J]. Acta Academiae Medicinae Qingdao Universitatis, 2010, 46(5): 377-379,383. DOI: 10.3969/j.issn.1672-4488.2010.05.001 | |
| Authors: | HE QING DU TING-TING SONG NING et al |
| Affiliation: | HE QING,DU TING-TING,SONG NING,et al(Department of Physiology,Shandong Provincial Key Laboratory of Pathogenesis and Prevention of Neurological Disorders and State Key Disciplines:Physiology,Qingdao University Medical College,Qingdao 266071,China) |
| Abstract: | Objective To construct the eukaryotic expression plasmid of human alpha-synuclein gene and detect its expression in human SK-N-SH neuroblastoma cells and its effect on cell viability.Methods PCR was performed to amplify the entire fragment of human alpha-synuclein gene,and the amplified gene was inserted into the eukaryotic expression vector pcDNA3.1(+)to form the new construct donated pcDNA3.1(+)/alpha-synuclein after purification.Liposome-mediated gene transfection method was used to transfect the human S... |
| Keywords: | alpha-synuclein plasmids Parkinson disease |
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