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].,2012,12(16):3068-3070 |
重叠PCR 法构建XLRS1 三种不同类型突变体 |
Construction of Three Different Types of Mutants of XLRS1 byOverlapping PCR |
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DOI: |
中文关键词: 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 |
英文关键词: XLRS1 Overlapping PCR Sited directed mutagenesis Retinoschisis |
基金项目:国家自然科学基金(30973257;81170855;81070743) |
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中文摘要: |
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。 |
英文摘要: |
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
ofNCBI. 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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