利用RACE(rapid-amplification of cDNA end)方法,以大豆叶片提取总的RNA为模板克隆了大豆Na+/H+逆向转运蛋白(Glycine max Na+/H+antiporter,GmNHX)基因,并将其连接到表达载体PBI121中,构建重组表达载体PBI121-NHX3.分析结果表明:该基...利用RACE(rapid-amplification of cDNA end)方法,以大豆叶片提取总的RNA为模板克隆了大豆Na+/H+逆向转运蛋白(Glycine max Na+/H+antiporter,GmNHX)基因,并将其连接到表达载体PBI121中,构建重组表达载体PBI121-NHX3.分析结果表明:该基因的ORF为1 503 bp,推测编码501个氨基酸.与所选取的10种植物同类蛋白氨基酸序列进行对比,一致性为72%～94%,并具有真核生物单价阳离子(氢离子)反向转运蛋白典型的结构域,将该基因命名为GmNHX3,GenBank接收号为JN872904.通过PCR和酶切鉴定,PBI121-NHX3构建成功.展开更多
[Objective] Genetic Engineering technology was used to regulate the expression of PttGA20-oxidase gene thus restrained plant height growth and internode elongation for cultivating dwarfed plant.[Method] Based on the R...[Objective] Genetic Engineering technology was used to regulate the expression of PttGA20-oxidase gene thus restrained plant height growth and internode elongation for cultivating dwarfed plant.[Method] Based on the RNAi principle,the gene specific sequences of PttGA20-oxidase in the antisense and sense orientations interrupted by a gene sequence from GUS were cloned into a binary vector pBI121.The selection marker gene npt Ⅱ was replaced with bar gene to RNAi plasmid.[Result] After undergone different endonuclease restrictions,the constructed constraint vector released different segments whose sizes were similar to that of target segment,which demonstrated that the RNAi plasmid of PttGA20-oxidase gene was successfully constructed.[Conclusion] The experiment provided a new way for culturing dwarfed plant.展开更多
文摘利用RACE(rapid-amplification of cDNA end)方法,以大豆叶片提取总的RNA为模板克隆了大豆Na+/H+逆向转运蛋白(Glycine max Na+/H+antiporter,GmNHX)基因,并将其连接到表达载体PBI121中,构建重组表达载体PBI121-NHX3.分析结果表明:该基因的ORF为1 503 bp,推测编码501个氨基酸.与所选取的10种植物同类蛋白氨基酸序列进行对比,一致性为72%～94%,并具有真核生物单价阳离子(氢离子)反向转运蛋白典型的结构域,将该基因命名为GmNHX3,GenBank接收号为JN872904.通过PCR和酶切鉴定,PBI121-NHX3构建成功.
文摘[Objective] Genetic Engineering technology was used to regulate the expression of PttGA20-oxidase gene thus restrained plant height growth and internode elongation for cultivating dwarfed plant.[Method] Based on the RNAi principle,the gene specific sequences of PttGA20-oxidase in the antisense and sense orientations interrupted by a gene sequence from GUS were cloned into a binary vector pBI121.The selection marker gene npt Ⅱ was replaced with bar gene to RNAi plasmid.[Result] After undergone different endonuclease restrictions,the constructed constraint vector released different segments whose sizes were similar to that of target segment,which demonstrated that the RNAi plasmid of PttGA20-oxidase gene was successfully constructed.[Conclusion] The experiment provided a new way for culturing dwarfed plant.
