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甘蓝型油菜生物素羧基载体蛋白基因的克隆与结构分析
引用本文:戴晓峰,卢长明,吴刚,武玉花,肖玲.甘蓝型油菜生物素羧基载体蛋白基因的克隆与结构分析[J].中国农业科学,2007,40(9):1883-1889.
作者姓名:戴晓峰  卢长明  吴刚  武玉花  肖玲
作者单位:中国农业科学院油料作物研究所,武汉,430062
基金项目:国家自然科学基金;国家重点基础研究发展计划(973计划);国家高技术研究发展计划(863计划)
摘    要: 【目的】生物素羧基载体蛋白负责将羧基从生物素羧化酶转移到羧基转移酶上,对于长链脂肪酸的从头合成与种子含油量的形成具有十分重要的作用。本研究的目的是从甘蓝型油菜品种基因组DNA中分离生物素羧基载体蛋白(BCCP)基因的全长编码区序列,通过生物信息学分析揭示该基因的结构特征(包括内含子的数量、长度与分布,蛋白质保守区疏水性特征等)和功能关系。【方法】基因克隆采取同源序列法进行。【结果】分离得到的bccp基因全长均为1 600 bp左右,根据其结构特点可以分成3种类型,它们均包含5个内含子。bccp1基因编码的蛋白质具备完整的生物素化功能结构域,是唯一功能完整的基因序列;bccp2和bccp3由于移码突变造成蛋白合成提前终止,翻译的蛋白质均缺少生物素化结构域。【结论】本研究首次从中国冬油菜品种中克隆获得bccp基因的全长序列并揭示了其序列特征,为进一步利用该基因开展含油量的基因调控研究提供了科学依据。

关 键 词:脂肪酸合成  生物素羧基载体蛋白(BCCP)  基因克隆  基因结构分析  甘蓝型油菜
收稿时间:2006-5-31
修稿时间:2006-05-31

Gene Cloning and Structural Analysis of Biotin Carboxyl Carrier Protein Encoding Gene in Brassica napus
DAI Xiao-feng,LU Chang-ming,WU Gang,WU Yu-hua,XIAO Ling.Gene Cloning and Structural Analysis of Biotin Carboxyl Carrier Protein Encoding Gene in Brassica napus[J].Scientia Agricultura Sinica,2007,40(9):1883-1889.
Authors:DAI Xiao-feng  LU Chang-ming  WU Gang  WU Yu-hua  XIAO Ling
Institution:Oil Crops Research Institute,Chinese Academy of Agricultural Sciences,Wuhan 430062
Abstract:Gene Cloning and Structural Analysis of Biotin Carboxyl Carrier Protein Encoding Gene in Brassica napus DAI Xiao-feng, LU Chang-ming, WU Gang, WU Yu-hua, Xiao Ling (Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, Hubei, China, 430062) Abstract: Acyl-CoA carboxylase (ACCase) catalyzes the first step in the de novo biosynthesis of fatty acids. Biotin carboxyl carrier protein (BCCP), being one of the subunits of plastidial ACCase, controls the transferring of carboxyl from acetyl-CoA carboxylase to carboxyltransferase. In this study, we isolated bccp genes from genomic DNA of three winter-type cultivars of Brassica napus and analyzed their structural aspects of these genes in both DNA and protein levels, including length, intron distribution, hydrophobic conserved regions etc.. Results showed that there were at least 3 isoforms of bccp genes found in genomes of B.napus. Only one isoform bccp1 encodes a protein which is full in length and contains the domain for biotinylation, while the other two isoforms bccp2 and bccp3 encode proteins that are incomplete and lack of the domain for biotinylation. Further analysis revealed that both in bccp2 and bccp3 an adenine insertion resulted in a shift-frame mutation. This research provides scientific basis of bccp genes for genetic engineering and molecular marker-assisstant breeding.
Keywords:De Novo fatty acid biosynthesis  Biotin carboxyl carrier protein (BCCP)  Gene cloning  Gene structure analysis  Brassica napus
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