sgna基因小麦高效表达载体的构建

Construction of high-level expression vectors containing sgna gene for use in wheat Transformation

利用在禾谷类作物中表达效率较高的启动子 Ubi对含目的基因 sgna的现有载体进行了改造 ,并引入筛选标记基因 bar;为提高目的基因的表达水平 ,在目的基因 5′端引入了Ω和 kozak序列 ,3′端引入了 poly( A)序列 ,成功构建了适用于小麦的抗虫基因植物表达载体p GU4AGBar和 p GBIU4AGBar。其中 p GU4AGBar含有顺向连接的 Ubi- sgna及 Ubi- bar基因表达盒 ,p GBIU4AGBar含有 T- DNA边界序列 ,并在其左右边界中间插入了含有顺向连接的 Ca MV35 S- npt 、Ubi- sgna和 Ubi- bar基因表达盒。

In the report,the maize ubiquitin 1 promoter Ubi ,a high efficiency promoter in gramineous crop,had been introduced into the original plasmid with s gna gene.Expression box of bar gene had been also inserted as a selectable marker.In order to improve the expression level of the target genes,Ω kozak sequence and poly (A) sequence were added to the 5′ end and 3′ end of the target genes,respectively.The wheat expression vectors with insect resisting gene pGU4AGBar and pGBIU4AGBar had been constructed,respectively.pGU4AGBar contains isodirectional expression box of Ubi s gna and Ubi bar .pGBIU4AGBar contains T DNA boundary sequence,and isodirectional expression box of CaMV35S nptⅡ,Ubi s gna and Ubi bar had been inserted within the T DNA boundary sequence.The sgna gene,a synthetical agglutinin gene of Galanthus navies,codes a poisonous protein for Homoptera .