[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.%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 DH5a in Genbank. Ge-nomic DNA of E. coli DH 5 a 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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