摘要
Growth Regulating Factor (GRF)家族基因是一类植物特有的转录因子,广泛参与植物多个重要的发育过程。为了研究大豆中的GRF转录因子的功能,本研究从大豆‘天隆1号’品种中克隆获得全长为1 038 bp的cDNA序列,由于该序列的编码蛋白与拟南芥GRF5具有高度的相似性,因此将其命名为GmGRF5基因。通过RT-PCR进行组织部位表达分析,发现GmGRF5基因在大豆各个组织部位均有表达,且在花中的表达量最高。通过ExPASy-Protparam分析发现,该蛋白质的分子量为39.463 48 kD,是具有一定亲水性、不稳定的碱性蛋白。该蛋白含有GRF家族蛋白的两个保守功能域(QLQ和WRC)。利用SOPMA对该蛋白的二级结构预测,显示其主要由无规则卷曲(72.05%)、α-螺旋(13.83%)、延伸链区(9.8%)和β-转角(4.32%)组成。此外,还利用不同的生物信息学软件对其三级结构、跨膜域、亚细胞定位、互作蛋白进行了分析,并构建系统进化树。结果表明,Gm GRF5基因在进化过程中具有高度的保守性,在大豆中具有潜在的功能多样性。
Growth Regulating Factor(GRF) family genes are a class of plant-specific transcription factors that are widely involved in many important developmental processes of plants. This study investigated the function of GRF transcription factors in soybean. A 1 038 bp cDNA sequence was cloned from soybean(Glycine max(L.) Merr.)cultivar ’Tianlong1’. Due to high sequence similarity to GRF5 protein in Arabidopsis, it was named as GmGRF5. The tissue expression pattern analyzed by real-time quantitative PCR revealed that GmGRF5 expressed in various tissues with the highest level in flowers. Analysis with ExPASy-Protparam showed that GmGRF5 encodes an alkaline protein of 345 amino acids with a molecular weight of 39. 46 348 kD and has hydrophilic instability. This protein contains two conserved domains of GRF family: QLQ and WRC. SOPMA was used to predict the secondary structure, which demonstrated that GRF protein consisted mainly of random coil(72.05%), α-helix(13.83%), extended chain region(9.8%), and β-turn(4.32%). In addition, different bioinformatics tools were employed not only to analyze its tertiary structure, transmembrane domain, subcellular localization, and protein interaction, but also to construct a phylogenetic tree. The results indicated that GmGRF5 is an evolutionally conserved protein and has diverse functionsin soybean.
作者
徐思远
刘志雄
张晓玫
Xu Siyuan;Liu Zhixiong;Zhang Xiaomei(College of Horticulture and Gardening,Yangtze University,Jingzhou,434023;MOA Key Lab of Soybean Biology(Beijing),National Key Facility for Crop Gene Resources and Genetic Improvement,Institute of Crop Science,Chinese Academy of Agricultural Sciences,Beijing,100081)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第5期1393-1400,共8页
Molecular Plant Breeding
基金
国家自然科学基金项目(31771714)
转基因生物新品种培育重大专项(2016ZX08004-005,2018ZX08009-24B)共同资助。
