深黄被孢霉Δ6脂肪酸脱氢酶基因的原核表达和转基因植株再生研究


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深黄被孢霉Δ6脂肪酸脱氢酶基因的原核表达和转基因植株再生研究

深黄被孢霉Δ6脂肪酸脱氢酶基因的原核表达和转基因植株再生研究

作者：王广科, 李明春, 李晋川, 刑来君

【摘要】 目的通过农杆菌介导法将来源于深黄被孢霉的Δ6脂肪酸脱氢酶(D6D)基因导入到东北大豆绥农10和吉林47中，最终获得转基因植株。方法利用子叶节外植体诱导出丛生芽和再生植株，利用卡那霉素的抗性筛选培养基连续筛选以及PCR方法、DNA分子杂交分析和GC分析检测转基因植株中的Δ6脂肪酸脱氢酶基因表达状况。结果经过含50 mg/L卡那霉素的抗性筛选培养基连续筛选，获得了一批转基因植株。经PCR检测和DNA分子杂交分析，初步证明Δ6脂肪酸脱氢酶基因已导入并整合到大豆基因组中。将转基因大豆T1代和T2代种子进行脂肪酸GC分析，结果在大豆种子中检测到Δ6脂肪酸脱氢酶基因的产物GLA，证明该基因在大豆中得到了表达。

【关键词】 Δ6脂肪酸脱氢酶基因；大豆；农杆菌介导转化；脂肪酸GC分析；转基因植株

【Abstract】 Objective The Δ6fatty acid desaturase genes from Mortitieralla isabellina were introduced into suinong10 and jilin47 via Agrobacterium infection method, some transgenic soybean were obtained. Methods Adentitious buds and regenerated plants were inducted from cotylendon nodes, Kanamycin resistant culture medium, PCR Southern blot and GC analysis detected condition of the Δ6fatty acid desaturase genes of transgenic plants. Result Kanamycin resistant(Kmr) transgenic plants were obtained by screening in selective condition. PCR and Southern blot analysis from transgenic plants proved that the Δ6fatty acid desaturase genes were transferred and integrated into soybean genomes. 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. The presence of GLA indicated that the Δ6fatty acid desaturase genes from Mortitieralla isabellina were expressed in transgenic soybeans.

【Key words】 △6fatty acid desaturase gene;Soybean(Glycin max L.);Agrobacteriummediated transformation;GC analysis of fatty acid;transgenic plant

GLA是多烯不饱和脂肪酸1（Polyunsaturated Fatty Acid，PuFA）n6代谢路线中的一种重要的脂肪酸［1］，它作为人体必需脂肪酸（Essential fatty acid，EFAs），对人体具

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