本文選題：油葵 + 脂肪酸去飽和酶��； 參考：《江蘇農(nóng)業(yè)學(xué)報(bào)》2017年02期

【摘要】：為研究油葵FAD2-1基因在脂肪酸合成中的催化功能,利用RT-PCR技術(shù),從油葵品種新葵雜4號未成熟種子中克隆HaFAD2-1基因的全長cDNA序列。將該基因序列構(gòu)建穿梭表達(dá)載體pYES2-HaFAD2-1,并轉(zhuǎn)化到營養(yǎng)缺陷型釀酒酵母菌株INVSc1中。將重組酵母誘導(dǎo)培養(yǎng)后對其總脂肪酸進(jìn)行GC-MS分析。結(jié)果顯示,HaFAD2-1基因編碼一個(gè)長378個(gè)氨基酸的脂肪酸去飽和酶蛋白,分子質(zhì)量43 719,等電點(diǎn)(pI)為8.38,具有3個(gè)高度保守的組氨酸簇。脂肪酸甲酯氣相色譜分析結(jié)果表明HaFAD2-1基因在釀酒酵母中獲得表達(dá),能將油酸轉(zhuǎn)化為亞油酸,證明克隆得到的HaFAD2-1具有完整的催化功能。
[Abstract]:In order to study the catalytic function of oil sunflower FAD2-1 gene in fatty acid synthesis, the full-length cDNA sequence of HaFAD2-1 gene was cloned from immature seeds of oil sunflower variety Xinkuiza 4 by RT-PCR technique. The shuttle expression vector pYES2-HaFAD2-1 was constructed and transformed into nutrition-deficient Saccharomyces cerevisiae strain INVSc1. The total fatty acids of recombinant yeast were analyzed by GC-MS after induction and culture. The results showed that HaFAD2-1 gene encodes a 378 amino acid long fatty acid desaturase protein with molecular weight of 43.719 and isoelectric point Pi of 8.38, with three highly conserved histidine clusters. Gas chromatographic analysis of fatty acid methyl ester showed that the HaFAD2-1 gene was expressed in Saccharomyces cerevisiae and could transform oleic acid into linoleic acid, which proved that the cloned HaFAD2-1 had complete catalytic function.
【作者單位】： 石河子大學(xué)生命科學(xué)學(xué)院/農(nóng)業(yè)生物技術(shù)重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金項(xiàng)目(31360052) 兵團(tuán)博士基金項(xiàng)目(2012BB005)
【分類號】：S565.5

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