GhTLP1基因特征分析、cDNA克隆及组织表达特性  

Feature Analysis, cDNA Cloning and Tissue Expression Characteristics of GhTLP1 Gene

作　　者：孙会忠[1] 杨月琴[1] 洪亚平[1] 戴攀峰[1] Sun Huizhong;Yang Yueqin;Hong Yaping;Dai Panfeng(College of Agriculture, Henan University of Science and Technology, Luoyang, 471003)

机构地区：[1]河南科技大学农学院

出　　处：《分子植物育种》2019年第10期3165-3170,共6页Molecular Plant Breeding

基　　金：河南省科技厅农业攻关计划项目(172102110020);河南省高等学校重点科研项目(16A180003;15A180003)共同资助

摘　　要：为了获取和验证类甜蛋白功能基因,并了解其基本的理化性质和组织表达特性,本研究在前期的海岛棉转录组高通量测序基础上,得到一个类甜蛋白功能基因的完整开放阅读框,命名为GhTLP1,通过全长特异引物,对GhTLP1进行了T-A克隆,测序结果符合已知序列。对GhTLP1基因序列特征分析结果表明,GhTLP1基因ORF序列全长为912 bp,共编码303个氨基酸残基,分子量为32.18 kD,等电点为4.36,第1~22位存在信号肽,无跨膜结构,预测属于亲水性蛋白;GhTLP1基因与其它植物的TLP基因的相似度达到50.15,具有完整的16个保守的半胱氨酸残基位点;GhTLP1基因与同源序列的聚类分析结果表明,GhTLP1基因与Gossypium hirsutum (XM016846902.1)和Gossypium arboreum (XM017769653.1)的TLP基因聚在同一分支上。本研究结果支持GhTLP1归为类甜蛋白基因的定性和功能判断。RT-PCR试验表明,GhTLP1在正常植株和受病原胁迫植株的根、茎、叶中均有表达,相对表达量顺序为根>叶>茎。GhTLP1可作为棉花抗病育种和材料筛选的调控靶标基因。In order to obtain and verify the functio nal genes of thaumatin-like protein, and clarify its basic physicochemical properties and tissue expression characteristics, this study obtained a complete open reading frame of a functional gene of thaumatin-like protein based on the previous high throughput sequencing of Gossypium barbadense, which was named as GhTLP1. T-A cloning was performed on GhTLP1 by full length specific primers, and the sequencing results were consistent with the known sequence. The sequence characteristic analysis of the GhTLP1 gene showed that the total length of the GhTLP1 gene ORF sequence was 912 bp, encoding a total of 303 amino acid residues. The molecular weight was 32.18 kD, and the isoelectric point was 4.36. Signal peptides were existed on the 1~22 sites. The protein was predicted to be hydrophilic without transmembrane structure. The similarity between the GhTLP1 gene and the TLP gene of other plants reached 50.15, with 16 intact conserved cysteine residues. The cluster analysis of GhTLP1 gene and homologous sequences showed that the GhTLP1 gene was clustered on the same branch with the TLP gene of Gossypium hirsutum(XM016846902.1)and Gossypium arboreum(XM017769653.1). The above results supported the qualitative and functional judgement of GhTLP1 as a thaumatin-like protein gene. RT-PCR test showed that GhTLP1 was expressed in roots, stems and leaves of normal plants and pathogeny stressed plants, and the relative expression level was root >leaf >stem.GhTLP1 could be used as a target gene for cotton disease resistance breeding and material selection.

关 键 词：海岛棉 类甜蛋白 特征分析 克隆 组织表达 

分 类 号：Q943.2[生物学—植物学] S432.2[农业科学—植物病理学]