海岛棉耐盐相关基因GbPLA1-32的克隆及生物信息学分析
Cloning and Bioinformatics Analysis of the Salt Tolerance Related Gene GbPLA 1-32 in Gossypium barbadense
张红 1张悦新 2徐楠 2芮存 2范亚朋 2王晶 2韩明格 2王琴琴 2孙亮庆 2陈修贵 2陆许可 2王俊娟 2王德龙 2王帅 2郭丽雪 2赵兰杰 2陈超 2陈全家 1叶武威3
作者信息
- 1. 新疆农业大学农学院,教育部棉花工程研究中心,乌鲁木齐,830052
- 2. 中国农业科学院棉花研究所,棉花生物学国家重点实验室,郑州大学研究基地,农业部棉花遗传改良重点开放实验室,安阳,455000
- 3. 新疆农业大学农学院,教育部棉花工程研究中心,乌鲁木齐,830052;中国农业科学院棉花研究所,棉花生物学国家重点实验室,郑州大学研究基地,农业部棉花遗传改良重点开放实验室,安阳,455000
- 折叠
摘要
为了解海岛棉GbPLA1-32基因结构特征及功能,本研究以海岛棉'海7124'为实验材料,利用前期耐盐转录组测序结果筛选得到差异表达基因并进行克隆.通过生物信息学分析,获得'海7124'GbPLA1-32基因全长CDS,分析该基因编码蛋白的理化性质、鉴定其亲水性与疏水性,对磷酸化和糖基化位点、信号肽和跨膜区以及蛋白质结构进行预测,确定亚细胞定位并构建系统进化树.结果表明,该基因编码区全长1 530 bp,编码509个氨基酸,编码蛋白在磷脂酶A1家族,属α/β-水解酶超家族.它属于亲水性蛋白,不存在明显信号肽且不具有跨膜结构,亚细胞定位于叶绿体中.同时,α-螺旋、延伸链、β-转角和无规则卷曲这4个元件构成了GbPLA1-32蛋白的二级结构.进化树分析表明海岛棉与陆地棉亲缘关系最为接近,根据其编码蛋白将该基因命名为GbPLA1-32.RT-qPCR分析结果表明,GbPLA1-32基因能够对PEG和盐两种胁迫做出响应.本研究结果可为进一步阐明GbPLA1-32在海岛棉中的生物学功能提供参考.
Abstract
In order to understand the structural characteristics and functions of the sea island cotton GbPLA 1-32 gene.In this study,the'Hai7124'in the sea island cotton was used as the experimental material,and the differen-tially expressed genes were screened and cloned using the results of salt-tolerant transcriptome sequencing.Through bioinformatics analysis,the full-length CDS of'Hai7124'GbPLA 1-32 gene was obtained.Analyze the physical and chemical properties of the protein encoded by the gene.Identify its hydrophilicity and hydrophobici-ty.Prediction of phosphorylation and glycosylation sites,signal peptides and transmembrane regions,and protein structure.Determine subcellular location and construct a phylogenetic tree.The results showed that the coding re-gion of the gene was 1 530 bp in length,encoding 509 amino acids,and the encoded protein was in the phospholi-pase Al family,belonging to the α/β-hydrolase superfamily.It is a hydrophilic protein,there is no obvious signal peptide and no transmembrane structure,subcellular localization in the chloroplast.At the same time,the four ele-ments of α-helix,extended strand,β-turn and random coil constitute the secondary structure of the GbPLAl-32 protein.The phylogenetic tree analysis showed that the relationship between sea island cotton and upland cotton is the closest.The gene was named GbPLA 1-32 according to its coding protein.The results of RT-qPCR analysis showed that the GbPLA 1-32 gene can respond to salt and alkali stress.The results of this study can provide refer-ences for further elucidating the biological functions of GbPLA 1-32 in Gossypium barbadense.
关键词
海岛棉/磷脂酶A1/基因克隆/表达分析/生物信息学分析Key words
Gossypium barbadense/Phospholipase A1/Gene cloning/Expression analysis/Bioinformatics analysis引用本文复制引用
基金项目
国家自然科学基金青年基金(32001460)
出版年
2024
NETL
NSTL
万方数据