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| 重叠PCR一步法对三种淀粉合成酶融合基因的构建 | |
| 引用本文： | 陈国梁,张金文,王蒂,陈宗礼,杨波波. 重叠PCR一步法对三种淀粉合成酶融合基因的构建[J]. 甘肃农业大学学报, 2010, 45(3) |
| 作者姓名： | 陈国梁 张金文 王蒂 陈宗礼 杨波波 |
| 作者单位： | 1. 延安大学生命科学学院,陕西省区域生物资源保育与利用工程技术研究中心,陕西,延安,716000;甘肃农业大学农学院,甘肃,兰州,730070 2. 甘肃农业大学农学院,甘肃,兰州,730070 3. 延安大学生命科学学院,陕西省区域生物资源保育与利用工程技术研究中心,陕西,延安,716000 |
| 基金项目： | 国家自然科学基金项目，延安大学专项科研基金，延安大学大学生科技创新训练项目 |
| 摘 要： | 采用重叠PCR一步法对马铃薯块茎3种淀粉合成关键酶gbss、ssⅢ和ssⅡ的基因片段进行融合.结果表明:该方法在PCR反应体系中通过6个引物3个模板扩增得到1个含有马铃薯块茎3种淀粉合成关键酶gbss、ssⅢ和ssⅡ基因片段的融合基因gs3s2;与经典的重叠PCR方法相比,该方法不需对PCR产物进行回收,而只需一个PCR反应就可以构建得到融合基因,是一种快速、方便有效的构建融合基因的方法. |
| 关 键 词： | 重叠PCR 淀粉合成酶 融合基因 马铃薯 |
Construction of a fused gene from three kinds of starch synthases of potato (Solanum tuberosum L.)by using one-step overlap-PCR |
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| CHEN Guo-liang,ZHANG Jin-wen,WANG Di,CHEN Zong-li,YANG Bo-bo. Construction of a fused gene from three kinds of starch synthases of potato (Solanum tuberosum L.)by using one-step overlap-PCR[J]. Journal of Gansu Agricultural University, 2010, 45(3) | |
| Authors: | CHEN Guo-liang ZHANG Jin-wen WANG Di CHEN Zong-li YANG Bo-bo |
| Abstract: | The fusion gene gs3s2 which consists of three key enzymes of potato tubers starch synthesis gbss, ssⅢ and ssⅡgene fragments was amplified by one-step overlap PCR.Six primers and three templates were used in this PCR reaction system. Furthermore, inhomogeneity template DNA was added separately to explore whether these gene segments could be extended. Our result indicated that fusion gene could be obtained only once-PCR reaction by PCR-driven overlap extension method and could not extracting PCR products from agarose gel. Compared with the classical overlap PCR method, this method is a real celerity, highly efficient technique for gene splicing. |
| Keywords: | overlap polymerase chain reaction starch synthase fusion gene potato |
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