甘蓝型油菜Fad2与Fae1基因双干扰RNAi载体的构建  

RNAi Vector Construction of the Fad2 and Fae1 Genes of Rapeseed(Brassica napus)

作　　者：彭琦[1] 张源[1] 双桑[1] 魏然[1] 伍林涛[1] 丁昕[1] 阮颖[1] 刘春林[1] 

机构地区：[1]湖南农业大学作物基因工程省重点实验室,长沙410128

出　　处：《生物技术通报》2007年第5期163-166,共4页Biotechnology Bulletin

摘　　要：将来源于甘蓝型油菜XY15的Fad2与Fae1基因编码区,长度分别为349bp和426bp的两个保守序列片段连接成807bp的大片段。然后分别以正反向插入到pFGC5941干扰载体查耳酮合成酶内含子的两端,构建成RNAi载体。以期在菜籽中转录后能有效抑制Fad2与Fae1两个基因的表达。所构建的RNAi载体最终序列全长3kb,经限制性内切酶消化、PCR扩增验证和序列测定,证明目标序列与GenBank数据库中的碱基序列一致,并且为正反向插入到pFGC5941载体上。通过农杆菌介导的油菜转化,已获得油菜抗性再生植株144个。To construct a RNAi vector of Fad2 and Fae1 genes,807bp fragment,among these base pairs 349bp from Fad2 gene and 426 from Fae1 gene,was inserted into the both sides of Chalcone synthase intron of pFGC5941 in forward and reverse direction,respectively.After the tests of restriction endonuclease digestion,PCR amplification and sequencing,the results showed that the sequence inserted in pFGC5941 is identical with the sequence published in Genbank and the two 807bp fragments were inserted at expectant clone sites in opposite direction.

关 键 词：油菜 油酸脱氢酶 脂肪酸延长酶 RNAI 

分 类 号：S565.4[农业科学—作物学] Q943.2[生物学—植物学]