| 重叠PCR 法构建XLRS1 三种不同类型突变体 |
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| 引用本文： | 王峰程海霞袁冬青夏鑫刘庆淮.重叠PCR 法构建XLRS1 三种不同类型突变体[J].现代生物医学进展,2012,12(16):3068-3070. |
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| 作者姓名： | 王峰程海霞袁冬青夏鑫刘庆淮 |
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| 作者单位： | 南京医科大学第一附属医院 江苏南京210029 |
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| 基金项目： | 国家自然科学基金(30973257;81170855;81070743) |
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| 摘 要： | 目的:构建X连锁视网膜劈裂(XLRS)三种不同类型突变体(点突变、缺失突变、插入突变),以便进一步构建不同类型的突变细胞模型。方法:根据NCBI gene数据库中XLRS1基因的CDS设计三对不同类型的引物,使用重叠PCR法获得突变片段,然后克隆入载体pLJM1-EGFP。结果:经测序验证,成功构建了三种不同类型突变体。结论:使用重叠PCR法构建三种不同类型突变体,为进一步研究XLRS1不同类型突变的致病机制奠定了基础。
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| 关 键 词： | X染色体连锁视网膜劈裂蛋白1 重叠PCR 定点突变 视网膜劈裂 |
Construction of Three Different Types of Mutants of XLRS1 by
Overlapping PCR |
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| WANG Feng,CHENG Hai-xi,YUAN Dong-qing,XIA Xin,LIU Qing-huai.Construction of Three Different Types of Mutants of XLRS1 by
Overlapping PCR[J].Progress in Modern Biomedicine,2012,12(16):3068-3070. |
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| Authors: | WANG Feng CHENG Hai-xi YUAN Dong-qing XIA Xin LIU Qing-huai |
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| Institution: | (The First Affiliated Hospital with Nanjing Medical University,Nanjing,210029,China) |
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| Abstract: | Objective: To construct three different types of mutations of XLRS1 for further modeling different types of mutant cell lines.Methods: Three different pairs of mutation primers were designed according to the CDS sequence of XLRS1 from the gene database of NCBI.Then three mutant fragments were obtained by overlapping PCR,which were further cloned into the vector pLJM1-EGFP.Results: Three types of mutant plasmids were verified by sequencing.Conclusions: Construction of three different types of mutants by overlapping PCR provided the foundation for investigating the pathogenesis of different types of XLRS1 mutatnts. |
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| Keywords: | XLRS1 Overlapping PCR Sited directed mutagenesis Retinoschisis |
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