[1]何早柯,周茜萍,王梦瑶,等.油葵脂肪酸去饱和酶基因HaFAD2-1的克隆与功能鉴定[J].江苏农业学报,2017,(02):273-279.[doi:doi:10.3969/j.issn.1000-4440.2017.02.006]
　HE Zao-ke,ZHOU Xi-ping,WANG Meng-yao,et al.Cloning and functional identification of a fatty acid desaturase gene, HaFAD2-1, from oil sunflower[J].,2017,(02):273-279.[doi:doi:10.3969/j.issn.1000-4440.2017.02.006]
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油葵脂肪酸去饱和酶基因HaFAD2-1的克隆与功能鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年02期
页码:
273-279
栏目:
遗传育种·生理生化
出版日期:
2017-04-30

文章信息/Info

Title:
Cloning and functional identification of a fatty acid desaturase gene, HaFAD2-1, from oil sunflower
作者:
何早柯; 周茜萍; 王梦瑶; 朱新霞; 祝建波; 孙黎
(石河子大学生命科学学院/农业生物技术重点实验室，新疆石河子832000)
Author(s):
HE Zao-ke; ZHOU Xi-ping; WANG Meng-yao; ZHU Xin-xia; ZHU Jian-bo; SUN Li
(Key Lab of Agriculture Biotechnology/College of Life Science， Shihezi University， Shihezi 832000, China)
关键词:
油葵; 脂肪酸去饱和酶; 基因克隆; 酿酒酵母转化; 功能分析
Keywords:
oil sunflower; fatty acid desaturase; gene clone; transgenic Saccharomyces cerevisiae; function analysis
分类号:
S565.5;Q785
DOI:
doi:10.3969/j.issn.1000-4440.2017.02.006
文献标志码:
A
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:
To study the catalytic function of oil sunflower FAD2-1 gene in fatty acid synthesis, a full length cDNA of FAD2 named HaFAD2-1 was isolated from developing seeds of Helianthus annuus L. Xinza 4 by RT-PCR. The HaFAD2-1 gene encodes a-378aa oleic acid desaturase with predicted molecular weight of 43 719, isoelectric point (pI) value of 8.38 and three his-boxes. HaFAD2-1 gene was sub-cloned into the shuttle vector pYES2 to generate the recombinant vector pYES2-HaFAD2-1 which was transformed into a defective mutant INVSc1 strain of Saccharomyces cerevisiae for expression. The total fatty acids of recombinant yeast were analyzed by GC-MS after induced culture. The result showed that transgenic yeast with the vector pYES2-HaFAD2-1 could catalyze oleic acid into linoleic acid, indicating that HaFAD2-1 is a functional enzyme of FAD2.

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备注/Memo

备注/Memo:
收稿日期：2016-06-23 基金项目：国家自然科学基金项目(31360052)；兵团博士基金项目(2012BB005) 作者简介：何早柯(1992-），男，湖北麻城人，硕士研究生，主要从事植物分子生物学研究。(Tel) 18086041837； (E-mail) hzkshzu@163.com 通讯作者：孙黎，(Tel) 0993-2057207； (E-mail) sunlishz@126.com
更新日期/Last Update: 2017-05-02