2024年2月29日 星期四
十字花科植物FAE1 基因的克隆与功能验证
Cloning and function verification of FAE1 gene in some species of Brassicaceae
2013年 第22卷 第1期 页码[8-13]    下载全文[0.9MB]  
摘要

对十字花科(Brassicaceae)植物非洲芥菜(Brassica tournefortii Gouan)、埃塞俄比亚芥(B. carinata A. Braun)、短喙芥( B. elongata Ehrhart)、芝麻菜[Eruca vesicaria subsp. sativa ( Miller) Thellung]、野萝卜( Raphanus raphanistrum Linn.)、Crambe filiformis Jacq.、菥蓂(Thlaspi arvense Linn.)、臭荠[Coronopus didymus (Linn.) Smith]、荠[Capsella bursa-pastoris (Linn.) Medikus]和小花碎米荠(Cardamine parviflora Linn.)的FAE1 基因进行了克隆、序列比对及功能验证。结果显示:上述前6 种1 亚种的FAE1 基因长度均为1 521 bp,臭荠的FAE1 基因长度为1 517 bp,荠和小花碎米荠的FAE1 基因长度为1 518 bp,GenBank 登录号为JX898749—JX898758;它们的序列相似性较高,相似度达89%;对位排列矩阵长度1 521 bp,其中包含保守位点1 051 个(69. 1%)、变异位点470 个(30. 9%)和简约信息位点232 个(15. 3%);臭荠、荠和小花碎米荠的FAE1 序列在第132 位分别缺失3 个碱基,臭荠的FAE1 基因在第515 位缺失1 个碱基。虽然荠和小花碎米荠的FAE1 基因编码505 个氨基酸、臭荠的FAE1 基因仅编码186 个氨基酸、其他种类的FAE1 基因均编码506 个氨基酸,但它们的氨基酸序列相似度高达88. 9%;各种类的氨基酸序列存在151 个变异位点,其中有6 个变异位点与种子芥酸含量相关。Western blot 及气相色谱分析结果表明:各种类的FAE1 基因在酵母中均能表达出预期的蛋白产物;在臭荠和小花碎米荠FAE1 基因的转化酵母细胞中无芥酸积累,而在其他种类FAE1 基因的转化酵母细胞中均有芥酸积累;此外,除荠外的其他8 种1 亚种植物的种子芥酸含量与转化酵母细胞中的芥酸含量正相关。

Abstract

Cloning, sequence alignment and function verification of FAE1 gene from Brassicaceae species including Brassica tournefortii Gouan, B. carinata A. Braun, B. elongata Ehrhart, Eruca vesicaria subsp. sativa (Miller) Thellung, Raphanus raphanistrum Linn., Crambe filiformis Jacq., Thlaspi arvense Linn., Coronopus didymus ( Linn.) Smith, Capsella bursa-pastoris ( Linn.) Medikus and Cardamine parviflora Linn. were carried out. The results show that the length of FAE1 gene from the front six species and one subspecies about mentioned all is 1 521 bp, that from Coronopus didymus is 1 517 bp, and that from Capsella bursa-pastoris and Cardamine parviflora is 1 518 bp, their GenBank accession number is from JX898749 to JX898758. Their sequences have higher similarity with a degree of 89% similarity. In the 1 521 bp alignment matrix, there are 1 051 conserved sites (69. 1%), 470 variable sites (30. 9%) and 232 parsimony informative sites (15. 3%). Sequence of FAE1 gene from Coronopus didymus, Capsella bursa-pastoris and Cardamine parviflora deletes three bases in the 132nd position, respectively and that from Coronopus didymus also deletes one base in the 515th position. Although, FAE1 gene from Capsella bursa-pastoris and Cardamine parviflora encodes 505 amino acids, that from Coronopus didymus encodes only 186 amino acids, and that from other species all encodes 506 amino acids, but the similarity of these amino acid sequences is up to 88. 9%. There are 151 variable sites among amino acid sequences, in which, 6 variable sites are related with erucic acid content in seed. Analysis results of  Western blot and gas chromotography show that FAE1 gene from each species can express the expected protein product in transformed yeast cell. And there is no erucic acid accumulation in transformed yeast cell of FAE1 gene from Coronopus didymus and Cardamine parviflora, but there is erucic acid accumulation in all transformed yeast cells of FAE1 gene from the other species. In addition, except Capsella bursa-pastoris, there is a positive correlation in erucic acid content between seed and transformed yeast cell of FAE1 gene from other eight species and one subspecies.

关键词十字花科; FAE1 基因; 克隆; 功能验证; 芥酸含量
Key wordsBrassicaceae; FAE1 gene; cloning; function verification; erucic acid content
作者庞慧, 李莹, 李密密, 严琴琴, 杭悦宇, 孙小芹
所在单位江苏省•中国科学院植物研究所(南京中山植物园) 江苏省植物迁地保护重点实验室, 江苏南京210014
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基金项目江苏省自然科学基金资助项目(BK2010476); 江苏省植物迁地保护重点实验室开放基金项目(迁201202)