摘要
将从高山被孢霉中克隆的△6-脂肪酸脱氢酶基因与植物表达载体pBI121连接,构建了重组质粒pBMICL6,采用农杆菌介导的大豆子叶节转化系统成功地将该基因导入到栽培大豆吉林35、吉林43、吉林47、绥农10、绥农14和黑农37等品种中,获得了一批转基因植株。经PCR检测和Southern杂交分析,证明外源基因已导入并整合到大豆的基因组中。Northern杂交结果表明,该基因在转基因大豆的mRNA水平上获得表达。通过气相色谱(GC-MS)对大豆种子进行脂肪酸成分分析,发现转基因大豆产生了γ-亚麻酸,其含量最高可达25.165％。
γ-linolenic acid (GLA, C18:3 Δ6.9.12) is an essential fatty acid in human diet. Δ6-fatty acid desaturase(D6D) is the rate-limiting enzyme in production of GLA. It can convert from linoleic acid(LA, C18:2 Δ9.12) to GLA. The Δ6-fatty acid desaturase gene from Mortitieralla alpina was transferred into cultivars Jilin35, Jilin43, Jilin47, Suinong10, Suinong14 and Heinong37 soybean successfully with Agrobacterium mediated cotylendon node transformation system. PCR and Southern hybridization analysis on the transgenic plants has proved that exogenous gene has been transferred and integrated into soybean genome. The Northern hybridization analysis showed that the D6D gene of transgenic soybean was expressed at mRNA level. Total fatty acids extracted from the positive transgenic soybean were analyzed by gas chromatography (GC) of methyl esters to confirm the transgenic soybean containing a functional Δ6-fatty acid desaturase gene. Among the seeds tested, the highest content of GLA amounts to 25.165% of the total fatty acids. The presence of GLA indicates that the Δ6-fatty acid desaturase gene from Mortitieralla alpina were expressed in transgenic soybean.
出处
《中国农业科学》
CAS
CSCD
北大核心
2004年第8期1104-1109,共6页
Scientia Agricultura Sinica
基金
国家自然科学基金资助项目(30200176)
天津市自然科学基金重点资助项目(013802511)
关键词
高山被孢霉
△^6-脂肪酸脱氢酶
大豆
基因表达
Soybean
Δ6-fatty acid desaturase gene
γ-linolenic acid
Agrobacterium-mediated
Mortitieralla alpina
