油用牡丹“凤丹白”中FAD8基因的克隆范例.doc

前言洛阳理工学院毕业设计（论文）油用牡丹“凤丹白”中FAD8基因的克隆摘 要在日常生活中，人们的食用油大多为植物油，而植物油脂的营养价值高低主要取决于植物体内脂肪酸的成分和含有量，脂肪酸中脂肪酸脱氢酶FAD8具备有催化二烯脂肪酸转变为三烯脂肪酸的性能，是决定亚油酸转变为亚麻酸合成的关键酶。在促进人体健康方面，α-亚麻酸具有十分重要的作用，主要表现在对人体的发育及成长、免疫反应、血脂调节等方面。所以，要想在短期内得到亚麻酸从而提高植物食用油的营养价值，利用基因工程手段是最重要的途径。本实验以油用牡丹“凤丹白”开花10天、20天、30天的种子为材料，使种子去皮液氮冷冻研磨，提取种子中的RNA，消化之后进行RNA电泳，根据RNA电泳条带对RNA进行检测。根据购买的FAD8序列，设计引物反转录得到cDNA的第一条链，进一步进行扩增、电泳，把符合的电泳条带进行胶回收，最后连接到T载体上进行转化，将转化液在含有Amp的培养基上培养，长斑、挑斑，最后得到所要的克隆片段。该基因的成功克隆为进一步研究脂肪酸脱饱和酶奠定了一定的基础。关键词：油用牡丹，“凤丹白”，FAD8基因，克隆Cloning of FAD8 gene from peony Phoenix Dan White＂in oilABSTRACTThe composition and content of fatty acids in plants have a very important role in the production of oils and fats. Fatty acid dehydrogenase FAD8 is a key enzyme in the synthesis of linoleic acid, which catalyzes the conversion of fatty acid to three fatty acids, Alpha flax acid plays a very important role in promoting the health of the human body. In this experiment, the seed of＂Feng Dan Bai＂,which is used in oil ,is 10 days, 20 days, 30 days, the seeds were peeled by liquid nitrogen freezing and grinding, RNA was extracted and digested by RNA electrophoresis, the RNA was detected by RNA electrophoresis. FAD8 sequence according to the purchase, design primer for reverse transcription cDNA first stand, further amplification, electrophoresis, consistent with the electrophoretic bands of plastic recycling and finally connected the T vector transformation, will be transformed into liquid culture Amp culture medium in the presence of, long spot, picked the spot, finally get to the cloned fragment. The successful cloning of the gene lays a foundation for further research on fatty acid. KEY WORDS: Oil Peony，Feng Dan Bai，FAD8 gene，Clone，目 录 TOC \o 1-3 \u 前 言 PAGEREF _Toc452401362 \h 1第1章 文献综述 PAGEREF _Toc452401363 \h 21.1 油用牡丹“凤丹白”概述 PAGEREF _Toc452401364 \h 21.1.1 牡丹 PAGEREF _Toc452401365 \h 21.1.2 油用牡丹“凤丹白” PAGEREF _Toc452401366 \h 21.1.3 牡丹籽油 PAGEREF _Toc452401367 \h 31.2 脂肪酸脱氢酶（FAD） PAGEREF _Toc452401368 \h 41.2.1