浙江农业学报 ›› 2023, Vol. 35 ›› Issue (4): 831-840.DOI: 10.3969/j.issn.1004-1524.2023.04.10

• 园艺科学 • 上一篇    下一篇

芜菁BrrLOX7基因克隆、表达及生物信息学分析

燕存尧(), 贾凯, 闫会转(), 高杰   

  1. 新疆农业大学 园艺学院,新疆 乌鲁木齐 830052
  • 收稿日期:2021-12-13 出版日期:2023-04-25 发布日期:2023-05-05
  • 通讯作者: *闫会转,E-mail: hzhyan1118@163.com
  • 作者简介:燕存尧(1997—),男,新疆昌吉人,硕士研究生,主要从事蔬菜栽培与生理研究。E-mail: 1033146390@qq.com
  • 基金资助:
    农业农村部园艺作物生物学与种质创制综合性重点实验室开放课题(IVF201701);新疆维吾尔自治区高校科研计划自然科学项目(XJEDU2017S017);新疆维吾尔自治区园艺学重点学科基金(2016-10758-3)

Cloning, expression and bioinformatics analysis of BrrLOX7 gene in turnip

YAN Cunyao(), JIA Kai, YAN Huizhuan(), GAO Jie   

  1. College of Horticulture, Xinjiang Agricultural University, Urumqi 830052, China
  • Received:2021-12-13 Online:2023-04-25 Published:2023-05-05

摘要:

脂氧合酶(lipoxygenase,EC 1.13.11.12;简称LOX),是一类由非血红素铁组成的脂肪酸双加氧酶,在植物的生长发育和环境胁迫响应中发挥着重要作用,本研究以4片真叶期芜菁幼苗为材料,利用同源克隆技术从芜菁中克隆BrrLOX7基因。利用生物信息学方法分析BrrLOX7基因序列。使用荧光定量PCR技术分析非生物胁迫下BrrLOX7基因表达模式。结果显示,克隆的BrrLOX7基因CDS长度为2 715 bp,编码904个氨基酸,编码的蛋白质分子量为103.10 ku,无跨膜结构和信号肽。亚细胞定位预测显示,该基因编码的蛋白位于叶绿体中,该蛋白与白菜和萝卜的序列同源性和遗传距离最近。不同胁迫下的表达模式显示,BrrLOX7基因参与对4种胁迫(盐胁迫、冷胁迫、热胁迫、干旱胁迫)的响应。研究结果将为BrrLOX7基因的功能研究奠定基础。

关键词: 芜菁, 脂氧合酶, 基因克隆, 生物信息学分析, 表达分析

Abstract:

Lipoxygenase (EC 1.13.11.12; LOX) is a class of fatty acid dioxygenases composed of non-heme iron, which plays an important role in plant growth and development and environmental stress response. In this study, the BrrLOX7 gene was cloned from turnip using homologous cloning techniques. The sequence of BrrLOX7 gene was analyzed by bioinformatics. The expression pattern of BrrLOX7 gene under abiotic stress was analyzed using fluorescence quantitative PCR. The results showed that the CDS length of cloned BrrLOX7 gene was 2 715 bp, it encoded 904 amino acids, protein encoded by BrrLOX7 gene had a relative molecular weight of 103.10 ku and had no transmembrane structure or signal peptide. Subcellular localization prediction showed that the protein encoded by this gene was located in chloroplasts, and the protein had the closest sequence homology and genetic distance to Chinese cabbage and radish. Expression patterns under different stresses showed that the BrrLOX7 gene was involved in response to four stresses (salt stress, cold stress, heat stress and drought stress). The results of this study will provide a basis for functional studies of the BrrLOX7 gene.

Key words: turnip, lipoxygenase, gene cloning, bioinformatics analysis, expression analysis

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