甘蓝型油菜及近缘种中异质型ACCase亚基accD编码基因的克隆及序列分析  

Cloning and Sequence Analysis of Heteromeric ACCase Subunit Encoding Gene accD from Four Close-related Species of Brassica

作　　者：王伏林[1] 吴关庭[1] 郎春秀[1] 刘仁虎[1] 陈锦清[1] 

机构地区：[1]浙江省农业科学院病毒学与生物技术研究所,杭州310021

出　　处：《农业生物技术学报》2008年第4期622-627,共6页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.30430450和No.30370910);浙江省自然科学基金项目(No.Y305251和No.M303144)资助。

摘　　要：采用RT-PCR方法,以甘蓝型油菜(Brassica napus)、甘蓝(B.oleracea)、芥菜型油菜(B.juncea)和白菜型油菜(B.rapa)的幼嫩叶片cDNA为模板对异质型乙酰辅酶A羧化酶(ACCase)的β-CT亚基(羧基转移酶)的accD编码基因进行扩增,得到长度分别为1470、1470、1464和1464 bp的编码序列,分别可编码489、489、487和487个氨基酸的蛋白质。白菜型油菜、芥菜型油菜和甘蓝的accD序列Genbank登录号分别为EU410075、EU410076和EU410077。序列分析表明,来自4个油菜近缘种的accD基因高度同源,编码的蛋白质都具有相似的锌指结构和C-末端5个相同的基元,其中基元Ⅰ(GSMGSVVG)和基元Ⅱ(PLIIVCASGGARMQE)在所有植物及大肠杆菌(Escherichia coli)的β-CT亚基中普遍存在。Southern杂交显示该基因在甘蓝型油菜及其近缘种的基因组中呈单拷贝。The cDNA fragments of accD gene encoding for subunit β-carboxyltransferase of heteromeric Acetyl-CoA carboxylase （ACCase） were amplified from young leaves cDNA of Brassica napus, B. olercea, B. juncea and B. rapa by RT-PCR. The lengths of the amplified accD genes were 1470, 1 470, 1464 and 1464 bp, respectively. Sequence analysis showed that the cDNA sequences of the four accD genes shared high homology. The amino acid sequences deduced from the four accD genes contained the similar zinc finger structure and five same motifs on C-terminal regions, among which motif I （GSMGSWG） and motif Ⅱ（PLⅡVCASGGARMQ E） were present in β-carboxyltransferases of all plant species and Escherichia coll. Southern blotting analysis demonstrated that accD was a single copy gene in genomes of the four close-related species of Brassica.

关 键 词：甘蓝型油菜 近缘种 乙酰辅酶A羧化酶(ACCase) accD基因 克隆 

分 类 号：S188[农业科学—农业基础科学]