生物技术通报2024,Vol.40Issue(12) :93-101.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0465

辣椒CabZIP42的克隆及表达分析

Cloning and Expression Analysis of CabZIP42 Gene in Pepper

李白雪 李金玲 陈春林 杜清洁 李猛 王吉庆 马勇斌 肖怀娟
生物技术通报2024,Vol.40Issue(12) :93-101.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0465

辣椒CabZIP42的克隆及表达分析

Cloning and Expression Analysis of CabZIP42 Gene in Pepper

李白雪 1李金玲 1陈春林 1杜清洁 1李猛 1王吉庆 1马勇斌 2肖怀娟1
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作者信息

  • 1. 河南农业大学园艺学院,郑州 450002
  • 2. 郑州普罗现代农业有限公司,郑州 450002
  • 折叠

摘要

[目的]探究辣椒bZIP基因在响应非生物胁迫应答过程中的作用,为提高辣椒抗逆性提供基因资源.[方法]以辣椒'G7'为材料,基于前期转录组测序得到一个候选基因CabZIP42,并对其进行克隆,通过生物信息学的方法对其编码蛋白的分子特征进行分析,并利用RT-qPCR技术分析其在不同组织、不同胁迫处理下的表达模式.[结果]CabZIP42编码区全长1233 bp,编码410个氨基酸,预测分子量为44.85 kD,理论等电点为9.47;属于亲水蛋白,脂肪族氨基酸指数为61.88,热稳定性较高.CabZIP42蛋白中包含2个保守结构域,碱性氨基酸区域N-x9-R和亮氨酸拉链区x6-L-x6-L-x6-L.该蛋白二级结构以无规则卷曲为主,还含有少量的a-螺旋.系统进化分析显示,CabZIP42与番茄、马铃薯的蛋白进化关系最近.启动子预测分析表明,CabZIP42启动子上存在多种激素和胁迫相关的顺式作用元件.[结论]CabZIP42在辣椒叶片中的表达量最高,并且参与ABA信号转导,响应干旱、高温与高盐胁迫.

Abstract

[Objective]To explore the role of bZIP gene in response to abiotic stress would provide genetic resources for improving the stress resistance of peppers.[Method]A candidate gene CabZIP42 was obtained and cloned based on transcriptome sequencing in pepper 4 G7',the molecular characteristics of the encoded protein were analyzed by bioinformatics methods,and its expression patterns were analyzed by RT-qPCR in different tissues and under different stress treatments.[Result]The coding region of CabZIP42 was 1 233 bp in length,encoding 410 amino acids,with a predicted molecular weight of 44.85 kD and a theoretical isoelectric point of 9.47.The CabZIP42 protein contained two conserved domains,the basic amino acid region N-x9-R and the leucine zipper region x6-L-x6-L-x6-L.The secondary structure of the protein was dominated by random coils,and contained a small amount of α-helix.Phylogenetic analysis showed that CabZIP42 had the most recent protein evolutionary relationship with tomato and potato.Promoter prediction analysis showed that there were multiple hormonal and stress-related cis-acting elements on the CabZIP42 promoter.[Conclusion]The expression of CabZIP42 gene was the highest in pepper leaves,and it was involved in ABA signal transduction in response to drought,high temperature and high salt stress.

关键词

辣椒/bZIP转录因子/克隆/表达分析/非生物胁迫

Key words

pepper/bZIP transcription factor/cloning/expression analysis/abiotic stress

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出版年

2024
生物技术通报
中国农业科学院农业信息研究所

生物技术通报

CSTPCD北大核心
影响因子:0.505
ISSN:1002-5464
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