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1、多聚磷酸盐激酶（ PPK ）基因植物表达载体的构建曹访;杨志红 ;韩志萍 ;杨倩 ;费佳玲【期刊名称】农业科学与技术(英文版) 【年(卷),期】2012(013)010【摘要】 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(L03
2、719) of E.coli DH5 a in Genbank.GenomicDNA ofE.coli DH 5 a was extracted as template for the amplificationof PPKgene by PCR method.By using In-Fusion HD Cloning Kit,the PPK gene was directionally cloned into NcoI site of the pCAMBIA1302 vector.Result Sequencing results showed that the 2.0 kb long fr
3、agment 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.%目的构建融合表达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 PPK基因序列(L03719 )设计引物，以E.coli DH5 a基因组DNA为模板，通过PCR扩增得PPK基因撚后用In-FusionHD Cloning Kit将PPK基因克隆到pCAMBIA1302 载体的Ncol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 基因已经插入植物表达载体 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-