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| 实时荧光定量聚合酶链反应突触后密度蛋白93基因TaqMan探针的制备 | |
| 引用本文： | 缪爱芳,陈梦玲,沈爱国,严美娟,高尚锋,程纯.实时荧光定量聚合酶链反应突触后密度蛋白93基因TaqMan探针的制备[J].南通医学院学报,2006,26(3):157-159. |
| 作者姓名： | 缪爱芳 陈梦玲 沈爱国 严美娟 高尚锋 程纯 |
| 作者单位： | 1. 南通大学继续教育学院,南通,226001 2. 南通大学基础医学院微生物与免疫学教研室,南通,226001 3. 南通大学基础医学院神经再生重点实验室,南通,226001 |
| 基金项目： | 中国科学院资助项目;国家自然科学基金;江苏省高校自然科学基金;江苏省社会发展基金;江苏省六大人才高峰基金 |
| 摘 要： | 目的:制备实时荧光定量聚合酶链反应(FQ-PCR)突触后密度蛋白93(PSD93)基因TaqM an探针以进一步研究PSD93表达情况。方法:采用RT-PCR法,从生后1天的SD大鼠大脑组织mRNA中扩增PSD93基因部分CDS区片段,克隆入T载体,筛选阳性克隆、酶切鉴定及序列测定,采用TaqM an探针,以重组质粒为模板绘制标准曲线。结果:RT-PCR法扩增出一特异产物与预期长度113bp相符,DNA序列测序的结果与GenBank提供的已知序列(RNU50717)比较,所克隆的PSD93基因片段与其中的113bp完全相同,与PSD93基因100%同源。以重组质粒为模板绘制标准曲线,相关系数r大于0.99,反应效率为0.95,各点变异系数小于20%。结论:采用RT-PCR和T载体技术成功获得FQ-PCR PSD93基因TaqM an探针。 |
| 关 键 词： | 聚合酶链反应 实时荧光定量 TaqMan探针 突触后密度蛋白93 大鼠 |
| 文章编号： | 1000-2057（2006）03-0157-03 |
| 收稿时间： | 2006-02-23 |
| 修稿时间： | 2006-02-23 |
The preparation of PSD93 TaqMan probe for real - time fluorescence quantitative PCR |
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| MIAO Aifang, CHEN Menghng, SHEN Atguo,et al.The preparation of PSD93 TaqMan probe for real - time fluorescence quantitative PCR[J].ACTA Academiae Medicinae Nantong,2006,26(3):157-159. | |
| Authors: | MIAO Aifang CHEN Menghng SHEN Atguo |
| Institution: | 1Department of Microbiology and Immunology, Basical Medical College, Nantong University, Nantong 226001 ; 2 Department of Continuing Education, Nantong University ;3The Key Laboratory of Neuroegeneration, Nantong Unirersity |
| Abstract: | Objective:To obtain the probe of postsynaptic density 93(PSD93) for real-time fluorescence quantitative PCR(FQ-PCR).Methods:The partial CDS of PSD93 was amplified by RT-PCR from 1d postnatal SD rat brain.RT-PCR product was ligated into pGEM-T vector,and the DNA sequence was detected.The standard curve was drawn with the recombinant plasmids and the PSD93 TaqMan probe.Results:The product of RT-PCR was 113 bp which matched the size of the purpose.This DNA sequence was 100% homogeneous with the PSD93 cDNA reported. The correlation coefficient of the standard curve was above 0.99,the reaction efficiency was 0.95 and all the variation coefficients were below 20%.Conclusion:The PSD93 TaqMan probe for FQ-PCR has been successfully obtained with RT-PCR and T vector techniques. |
| Keywords: | Polymerase Chain reaction Real-time fluorescence quantification TaqMan Probe Postsynaptic dlensity 93 Rats |
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