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当前位置: 首页 > 文章 > 莱茵衣藻ω-3脂肪酸脱氢酶基因在毕赤酵母中的表达与活性分析 生物技术通报 2011,0 (12) 96-101
Position: Home > Articles > Hetero Expression of ω-3 Desaturase Gene from Chlamydomonas reinhardtii and Assay of Its Activity in Pichia pastoris Biotechnology Bulletin 2011,0 (12) 96-101

莱茵衣藻ω-3脂肪酸脱氢酶基因在毕赤酵母中的表达与活性分析

作  者：
牛丽红;耿丽丽;张蕊;孙长坡;张杰
单  位：
国家粮食局科学研究院;中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室
关键词：
莱茵衣藻;ω-3脂肪酸脱氢酶;α-亚麻酸;毕赤酵母
摘  要：
采用RT-PCR方法克隆到莱茵衣藻ω-3脂肪酸脱氢酶基因lyd15,与毕赤酵母表达载体pPIC3.5K连接,电击法转化毕赤酵母GS115。转化子经高浓度G418筛选出高抗性重组子,经PCR鉴定目的基因已整合入毕赤酵母基因组中。甲醇诱导表达,RT-PCR检测表明莱茵衣藻ω-3脂肪酸脱氢酶基因在毕赤酵母中得到了表达;毕赤酵母总脂肪酸甲酯经气相色谱(GC)分析结果显示亚油酸的含量明显降低,而α-亚麻酸的含量有所提高。
译  名：
Hetero Expression of ω-3 Desaturase Gene from Chlamydomonas reinhardtii and Assay of Its Activity in Pichia pastoris
作  者：
Niu Lihong1 Geng Lili1 Zhang Rui2 Sun Changpo2 Zhang Jie1(1 State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193;2Academy of State Administration of Grain,Beijing 100037)
关键词：
Chlamydomonas reinhardtii ω-3 fatty acid desaturase Alpha-linolenic acid Pichia pastoris
摘  要：
The C.reinhardtii ω-3 fatty acid desaturase gene(lyd15) was subcloned into the methylotrophic yeast Pichia pastoris expression vector pPIC3.5K to generate a recombinant plasmid p3.5K-Ld3,which was subsequently transformed into Pichia pastoris strain GS115 for heterologous expression with electroporation method.The expression of desaturase gene lyd15 in P.pastoris was verified with RT-PCR.Total fatty acids were extracted from the induced cells and methylated.The fatty acid methyl esters were analyzed with gas chromatography(GC),the results show that the amount of linoleic acid was decreased remarkably,whereas the content of alpha-linolenic acid was increased.

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