| 摘要: |
| 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cry Ia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-cry Ia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cry Ia可以编码对鳞翅目昆虫具有毒杀作用的蛋白. |
| 关键词: 表达载体 启动子 基因 质粒 小麦 |
| DOI： |
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| 基金项目:国家“九五”重点科技攻关项目(专题)(96-009-01-14)；杨凌农业生物技术育种中心开放课题资助项目(1999-19) |
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| Construction of high-level expression vector based on cry Ia gene for use in wheat transformation |
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| Abstract: |
| 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'-endof 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. |
| Key words: expression vector promoter gene plasmid wheat |
