DU Jing-hong,LIU Li-jun.Construction of fad3c Gene Silencing Vector in Soybean[J].Soybean Science,2013,32(01):28-37.[doi:10.3969/j.issn.1000-9841.2013.01.007]
大豆fad3c基因沉默载体的构建
- Title:
- Construction of fad3c Gene Silencing Vector in Soybean
- 文章编号:
- 1000-9841(2013)01-0028-05
- 分类号:
- S565.1
- 摘要:
- 构建大豆fad3c基因的沉默载体,旨在为培育生物安全的高油酸低亚麻酸转基因大豆奠定基础。从高蛋白大豆黑农35中克隆大豆种子特异表达启动子GY1;从高油大豆黑农37中克隆fad2-1b基因内含子1;从5个转基因受体大豆中克隆fad3c基因片段,作为正向臂和反向臂。构建筛选标记基因为bar,启动子为GY1,内含子为fad2-1b基因内含子1的双T-DNA载体。将双T-DNA载体转化大肠杆菌DH5α,酶切和序列分析鉴定表明扩增得到的重组质粒正确,命名为pDT-GFAD3I。
- Abstract:
- Soybean fad3c gene silencing vector was constructed for breeding high oleic acid and low linolenic transgenic soybeans with biosafety.Seed-specific promoter GY1 of soybean was amplified from high protein soybean ‘Heinong 35’;fad2-1b gene intron 1 was amplified from high oil soybean ‘Heinong 37’;fad3c gene fragment was amplified from five transgenic acceptors of soybean,which was used as forward arm and reverse arms.These target fragments were cloned into the TA cloning vector with selection marker bar? gene,promoter GY1 and fad2-1b ?gene intron 1.Then the recombination plasmid was introduced into E.coli DH5α,and the plasmid was verified by enzymes digestion and sequence analysis.The recombination plasmid was named pDT-GFAD3I.
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备注/Memo
基金项目：黑龙江省农业科学院博士后科研启动经费。
第一作者简介： 杜景红（1968-），女，副教授，博士，主要从事水稻遗传育种研究和水稻抗逆性研究。E-mail：aimi933@163.com。
通讯作者： 刘丽君（1958-），女，研究员，主要从事大豆栽培育种研究。E-mail：nkyssbd@126.com。