作物学报2016，Vol.42Issue(7)：1000-1008,9.DOI:10.3724/SP.J.1006.2016.01000
甘蓝型油菜BnFAD2基因的克隆、表达及功能分析
Cloning and Expression Analyses forBnFAD2 Genes in Brassica napus
摘要
Abstract
The oil containing high oleic acid is high nutritional. InBrassica napus, the fatty acid desaturase gene (FAD2) is the key gene controlling oleic acid content. In this study, the full-length cDNA sequences of three genes located on chromosome A5, C5 and A1 inBrassica napus were cloned and namedBnFAD2-A5,BnFAD2-C5 andBnFAD2-A1. The three genes encode proteins with 384, 384, and 136 amino acid residues, respectively. TMHMM was used to predict transmembrane domain and Clust X soft-ware was used to analyze the activity center ofFAD2 genes. Both of the results showed thatBnFAD2-A1 did not have the function of dehydrogenase. The yeast complementary experiment on four genes (including publishedBnFAD2-C1gene) showed that the desaturation capability of BnFAD2-A5 gene was next to that ofBnFAD2-C5 gene, and both of them were greater than that of BnFAD2-C1 gene. The expression patterns of the four genes were analyzed by using qRT-PCR technique in different tissues and the protein stability of BnFAD2-C1, BnFAD2-A5, and BnFAD2-C5 was analyzed by using Hemagglutinin labeling method. Both of the results revealed that BnFAD2-A5 andBnFAD2-C5aremajor genes affecting the accumulation of oleic acid inB. napus.关键词
甘蓝型油菜/BnFAD2基因/生物信息学/蛋白活力/蛋白稳定性Key words
Brassica napus/BnFAD2/Bioinformation/Protein activity/Protein stability引用本文复制引用
刘睿洋,刘芳,官春云..甘蓝型油菜BnFAD2基因的克隆、表达及功能分析[J].作物学报,2016,42(7):1000-1008,9.基金项目
本研究由湖南省科技创新项目(CX2013A012)和国家重点基础研究发展计划(973计划)项目(2015CB150200)资助。 This study was supported by Innovation Foundation of Hunan Province (CX2013A012) and the National Basic Research Program of China (2015CB150200) （CX2013A012）
