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花生Δ¹²-脂肪酸去饱和酶基因RNAi表达载体的构建

RNAi Plasmid Construction of Delta-12-fatty Acid Desaturase Gene from Peanut

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【Author】 CHEN Zhan-kuan~1,ZHANG Xin-you~2,MIAO Li-juan~1,HUANG Bing-yan~1,TANG Feng-shou~2,ZHANG Zhong-xin~2(1.Biotechnology Research Institute,Henan Academy of Agricultural Sciences,Henan Key Laboratory for Crop Improvement,Zhengzhou 450002,China;2.Cotton and Oil Crops Research Institute,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)

【机构】河南省农业科学院生物技术研究所河南省农作物新品种重点实验室；河南省农业科学院棉花油料作物研究所；河南省农业科学院棉花油料作物研究所河南郑州450002；河南郑州450002；

【摘要】利用RNAi原理构建花生Δ12-脂肪酸去饱和酶基因的hpRNA表达载体,以抑制该基因的表达,获得高油酸/亚油酸比值的花生种质。根据花生Δ12-脂肪酸去饱和酶基因序列(GenBank:AF248739)设计引物,以豫花4号花生品种DNA为模板,克隆了该基因的启动子及长度约500 bp的外显子片段,在此基础上构建完成由自身启动子引导的花生Δ12-脂肪酸去饱和酶基因的反向重复片段RNAi载体pCAMBIA1301-Afad12Ri。更多还原

【Abstract】 An RNAi plasmid is constructed to inhibit the expression of delta-12-fatty desaturase gene in peanut,in order to down regulate the biosynthesis of linoleic acid and obtain transgenic lines with high oleic acid / linoleic acid ratio.The primers were designed based on the sequence of delta-12-fatty desaturase gene released in GenBank(AF248739),and the promoter region and partial extron of the gene in peanut was cloned using the genomic DNA from peanut cultivar Yuhua 4 as template.An RNAi plasmid pCAMBIA1301-Afad12Ri was constructed which contains the cloned promoter and inverted repeats of delta-12-fatty desaturase gene fragment.更多还原