生物技术通报 ›› 2013, Vol. 0 ›› Issue (10): 76-80.

• 研究报告 • 上一篇    下一篇

全雌系苦瓜ACC氧化酶基因cDNA克隆及序列分析

王日升1,2, 张曼1,黄如葵1,刘文君1,董文斌1,车江旅2, 方锋学1,李杨瑞2   

  1. (1. 广西农业科学院蔬菜研究所,南宁 530007 ;2. 广西作物遗传改良生物技术重点开放实验室,南宁 530007)
  • 收稿日期:2013-04-01 修回日期:2013-10-15 出版日期:2013-10-14 发布日期:2013-10-15
  • 作者简介:王日升, 男, 硕士, 副研究员, 研究方向: 蔬菜分子生物学;E-mail :shengriwang@126.com
  • 基金资助:

    广西自然科学基金重点项目(2010GXNSFD013031), 广西青年科学基金项目( 桂科自0991077), 第46 批教育部留学回国人员科研启动基金,“ 广西特聘专家” 专项经费

Cloning and Sequence Analysis of 1-aminocyclopropane-1-carboxylate (ACC)Oxidase Gene cDNA from Gynoecious Momordica charantia

Wang Risheng1,2,, Zhang Man1,Huang Rukui1,Liu Wenjun1,Dong Wenbin1,Che Jianglü2, Fang Fengxue1,Li Yangrui2,   

  1. (1. Vegetable Research Institute,Guangxi Academy of Agricultural Sciences,Nanning 530007 ;2. Guangxi Crop Genetic Improvement and Biotechnology Lab,Nanning 530007)
  • Received:2013-04-01 Revised:2013-10-15 Published:2013-10-14 Online:2013-10-15

摘要:

为研究1-氨基环丙烷-1-羧酸氧化酶(ACC 氧化酶,ACO) 基因在苦瓜性别分化中的作用,以全雌系苦瓜‘X-Hei-d-d’ 花蕾为试材,采用RT-PCR 和RACE 技术获得了ACC 氧化酶基因(Mc-ACO1) 的全长cDNA 序列。该序列为1 137 bp(GenBank 登录号:FJ459813),其完整开放阅读框长1 005 bp, 编码334 个氨基酸,预测分子量为37.30 kD, 肽链N 端缺失ACOs 家族典型的1 个保守区和2 个保守二价铁离子/ 抗坏血酸依赖型双加氧酶氨基酸残基。序列分析表明,植物ACO 基因的演化与其来源植物亲缘关系的一致性较高;与其他物种相比,苦瓜Mc-ACO1 不同的氨基酸序列主要表现在肽链N 端;苦瓜Mc-ACO1 应属于黄瓜Cs-ACO2 类成员,功能可能与性别分化相关。

关键词: 全雌系苦瓜, 性别分化, ACC 氧化酶, 基因克隆, 序列分析

Abstract:

The full cDNA sequence of 1-aminocyclopropane-1-carboxylic acid oxidase(ACO)gene was cloned by RT-PCR and RACE from floral buds of gynoecious bitter melon‘X-Hei-d-d’, which was named as Mc-ACO1(GenBank accession No. FJ459813). The full length cDNA of Mc-ACO1 is 1 137 bp, which includes 1 005 bp complete CDS and encodes 334 amino acid residues with a putative molecular mass of 37.30 kD. The protein encoded by Mc-ACO1 gene lacks one typically conserved domain and two amino acid residues of Fe2+ ascorbate family of dioxygenases. The bioinformatics characterization indicate that the evolution of plant ACO has higher consistence with their genetic relationship ; and the different amino acid sequences of Mc-ACO1 are mainly at the N-terminal area compared with those of other plants ;Mc-ACO1 should belong to the same member as cucumber Cs-ACO2, and the presumed function of Mc-ACO1 is probably related with sex differentiation.

Key words: Gynoecious bitter melon, Sex differentiation, ACC oxidase, Gene clone, Sequence analysis