cry Ia基因小麦高效表达载体的构建

Construction of high-level expression vector based on cry Ia 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cryIa的现有载体进行了改造,并引入了筛选标记基因bar,为提高目的基因的表达水平,还在目的基因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pGU4ABBar,该载体含有顺向连接的Ubi-cryIa及Ubi-bar基因表达盒,可用于基因枪、花粉管通道法等介导的小麦遗传转化,人工合成的苏云金芽孢杆菌杀虫蛋白基因cryIa可以编码对鳞翅目昆虫具有毒杀作用的蛋白。

In the report,the maize ubiquitin 1 promoter Ubi,a high efficiency promoter in gramineous crop,had been introduced into the original plasmid with cry Ia gene.Expression box of bar gene had also been 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 insectresisting gene wheat expression vector pGU4ABBar had been constructed.pGU4ABBar contains direct inserted expression box of Ubicry Ia and Ubibar.It is suitable for genetic transformation of wheat mediated by microprojectle or pollentube pathway.The cry Ia gene,a synthetical insecticidal crystal protein gene of Bacillus thuringiensis,may code a poisonous protein for Lepidoptera.