| 多聚磷酸盐激酶(PPK)基因植物表达载体的构建 |
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| 引用本文： | 曹访,杨志红,韩志萍,杨倩,费佳玲.多聚磷酸盐激酶(PPK)基因植物表达载体的构建[J].安徽农业科学,2012(31):15139-15141. |
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| 作者姓名： | 曹访 杨志红 韩志萍 杨倩 费佳玲 |
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| 作者单位： | 湖州师范学院,浙江湖州,313000 |
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| 基金项目： | 国家自然科学基金(31070451);浙江省钱江人才计划项目(2009R10016);浙江省自然科学基金(Y5110057) |
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| 摘 要： | 目的]枸建融合表达PPK和绿色荧光蛋白的融合表达载体pCAMBIA1302-PPK.方法]根据GenBank中登录的大肠杆菌PP基因序列(L03719)设计引物,以E.coli DH5α基因组DNA为模板,通过PCR扩增得PPK基因,然后用In-Fusion@HD Cloning Kit将PPK基因克隆到pCAMBIA1302载体的Nco Ⅰ酶切位点.结果]序列测定结果显示,pCAMBIA1302-PPK含有约2.0kb的PPK基因片段,说明PPK基因已插入植物表达载体pCAMBIA1302的绿色荧光蛋白基因前.结论]成功构建了融合表达PPK和绿色荧光蛋白的融合表达载体pCAMBIA1302-PPK.
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| 关 键 词： | 大肠杆菌 多聚磷酸盐激酶基因 植物表达载体 构建 |
Construction of Plant-based Expression Vector of Polyphosphate Kinase Gene |
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| Institution: | CAO Fang et al(Huzhou Teachers Collage,Huzhou,Zhejiang 313000) |
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| Abstract: | Objective] The aim was to construct the fusion expression vector of polyphosphate kinase(PPK) and green fluorescent protein(GFP) genes.Method] In this study,the primers were designed based on PPK gene sequence(L03719) of E.coli DH5α in Genbank.Genomic DNA of E.coli DH 5α was extracted as template for the amplification of PPK gene by PCR method.By using In-Fusion@ HD Cloning Kit,the PPK gene was directionally cloned into Nco I site of the pCAMBIA1302 vector.Result] Sequencing results showed that the 2.0 kb long fragment of PPK gene was inserted into the plant-based expression vector pCAMBIA1302 in front of GFP gene.Conclusion] The fusion expression vector of PPK and GFP genes were successfully constructed. |
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| Keywords: | Escherichia coli PPK gene Plant-based expression vector Construction |
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