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高山被孢霉△~6-脂肪酸脱氢酶基因转化大豆的研究

Introduction of △~6-fatty Acid Desaturase Gene from Mortieralla alpina into Soybeans by Agrobocterium Infection

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【作者】 卜云萍李明春胡国武邢来君

【Author】 BU Yun-ping,LI Ming-chun,HU Guo-wu,XING Lai-jun(Department of Microbiology,Nankai University,Tianjin 300071,P.R.China)

【机构】 南开大学微生物系南开大学微生物系 天津300071天津300071天津300071

【摘要】 采用农杆菌介导的大豆子叶节转化系统成功地将高山被孢霉△6-脂肪酸脱氢酶基因导入到栽培大豆吉林 4 3和黑农37品种中。经农杆菌浸染和共培养后 ,在含 5 0mg/L卡那霉素的选择培养基上连续筛选 ,获得一批转基因植株。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。通过GC -MS对大豆种子进行脂肪酸色谱分析 ,结果表明 ,产生了γ -亚麻酸 ,其含量最高可达 18.2 3%。

【Abstract】 The △6-fatty acid desaturase gene from Mortitieralla alpina was transferred into cultivars Jilin43 and heinong37 soybean successfully with Agrobacterium-cotylendon node transformation system.Cotyledon node explants were infected and co-cultivated with Agrobacterium tumefaciens.Kanamycin resistant (Kmr) plantlets were obtained by 3-4 cycles screening in selective condition.PCR and Southern hybridization analysis from the transferred gene plants proved that foreign gene were transferred and integrated into soybean genome.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 under detection,the highest content of GLA amounts to 18.23% 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.This is the first report at home and abroad so far about the expression of Mortitieralla alpina Δ6-fatty acid desaturase gene in soybean.

【基金】 国家自然科学基金项目 (30 2 0 0 1 76) ;天津市自然科学基金重点项目 (0 1 380 2 51 1 )
  • 【分类号】Q786;Q943.2
  • 【被引频次】14
  • 【下载频次】133
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