油葵脂肪酸去饱和酶基因HaFAD2-1的克隆与功能鉴定

发布时间：2018-05-16 23:24

本文选题：油葵 + 脂肪酸去饱和酶　；参考：《江苏农业学报》2017年02期

【摘要】：为研究油葵FAD2-1基因在脂肪酸合成中的催化功能,利用RT-PCR技术,从油葵品种新葵杂4号未成熟种子中克隆HaFAD2-1基因的全长cDNA序列。将该基因序列构建穿梭表达载体pYES2-HaFAD2-1,并转化到营养缺陷型酿酒酵母菌株INVSc1中。将重组酵母诱导培养后对其总脂肪酸进行GC-MS分析。结果显示,HaFAD2-1基因编码一个长378个氨基酸的脂肪酸去饱和酶蛋白,分子质量43 719,等电点(pI)为8.38,具有3个高度保守的组氨酸簇。脂肪酸甲酯气相色谱分析结果表明HaFAD2-1基因在酿酒酵母中获得表达,能将油酸转化为亚油酸,证明克隆得到的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.
【作者单位】：石河子大学生命科学学院/农业生物技术重点实验室;
【基金】：国家自然科学基金项目(31360052) 兵团博士基金项目(2012BB005)
【分类号】：S565.5

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