CaMV 35S增强子调节玉米基因表达的机制研究
Study on the Mechanism of CaMV 35S Enhancer Regulating Gene Expression in Maize (Zea mays)
郭金洁 1张丹 1张伟 1张方东 1杜邓襄2
作者信息
- 1. 华中农业大学植物科学技术学院,武汉430070
- 2. 武汉轻工大学生命科学技术学院,武汉430023
- 折叠
摘要
玉米(Zea mays)是重要的粮食作物、饲料来源和工业原料等,随着生物技术的发展,转基因等新型技术在玉米育种中的应用越来越广泛.花椰菜花叶病毒(Cauliflower mosaic virus,CaMV)启动子CaMV 35S及其增强子广泛应用于玉米等作物的转基因操作中,因此,研究CaMV 35S增强子的的作用机理对于今后的应用具有重要的理论价值.本研究从已经获得的玉米激活标记突变体库中筛选出突变体材料,在分离突变位点在染色体上分布的基础上,利用qPCR分析了转座子插入位点上游和下游约100 kb范围内基因的时空表达变化.结果表明,CaMV 35S增强子显著上调了邻近基因的表达(P<0.05),转录增加的基因分布在转座子插入位点的上下游,激活距离可达63.7 kb.在上调表达的4个基因中,基因表达水平增强呈剂量依赖性,纯合突变体的表达显著高于杂合突变体(P<0.05).CaMV 35S增强子虽然提高了玉米突变体基因的转录水平,但并未显著改变基因在不同组织中的表达模式.CaMV 35S增强剂在玉米中的作用范围、剂量效应和组织特异性为后续在玉米分子生物学和基因工程中的应用提供了实验依据,也为CaMV 35S启动子在其他植物中的应用提供了借鉴.
Abstract
Maize (Zea mays) is an important food crop, feed source and industrial raw material. With the development of bio-technology, transgenic and other new technologies are widely used in maize breeding. Cauliflower mosaic virus 35S (CaMV 35S) promoter and its enhancer are widely used in transgenic operation of maize and other crops. Therefore, studying the mechanism of CaMV 35S enhancer has important theoretical value for future application. Two trait mutants were screened from a maize activation-tagging mutant library previously constructed by the research group on genetic improvement of maize. Based on the identification of artificial transposon insertion sites in the chromosomes, the spatio-temporal variation of gene RNA expression in the range of about 100 kb upstream and downstream of the transposon insertion sites was analyzed using qPCR. The results showed that the enhancer of CaMV 35S significantly up-regulated the gene expression of adjacent genes (P<0.05), and the transcriptionally increased genes were distributed upstream and downstream of the transposon insertion site, with activation distance of up to 63.7 kb. Of the 4 genes up-regulated expression, the enhancement was dose-dependent, and the expression of the homozygous mutant was significantly higher than that of the heterozygous mutant (P<0.05). Although the CaMV 35S enhancer increased the transcription level of genes in the maize mutants, it did not significantly change the expression pattern of genes in different tissues. The scope, dosage effect, and tissue specificity of CaMV 35S enhancer in maize provide an experimental basis for subsequent use in maize molecular biology, and genetic engineering, and can also provide reference for the application of the promoter in other plants.
关键词
玉米/CaMV/35S增强子/CaMV/35S启动子/基因表达特征Key words
Maize/CaMV 35S enhancer/CaMV 35S promoter/Gene expression characteristics引用本文复制引用
基金项目
国家自然科学基金(31871636)
国家自然科学基金(31271737)
国家自然科学技术重大专项(2014ZX08003003)
国家自然科学技术重大专项(2016ZX08003003)
出版年
2023
国家科技期刊平台
NSTL
维普
万方数据