水滴伪康纤虫PpCaMK1基因克隆及生物学特征分析  

Cloning and Biological Characteristics of PpCaMK1 in Pseudocohnilembus persalinus

作　　者：吕艳红 曾红[1] 林童 林能锋[2] LYU Yanhong;ZENG Hong;LIN Tong;LIN Nengfeng(College of Life Sciences,Fujian Normal University,Fuzhou,Fujian 350002,China;Institute of Biotechnology,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350002,China)

机构地区：[1]福建师范大学生命科学学院,福建福州350002 [2]福建省农业科学院生物技术研究所,福建福州350002

出　　处：《福建农业学报》2025年第1期29-37,共9页Fujian Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2022YFD2401002)。

摘　　要：【目的】钙调素依赖蛋白激酶(calmodulin-dependent protein kinases,CaMKs)是钙离子介导的信号转导通路中的关键组成部分,可能成为治疗盾纤虫病的药物靶点。本研究试图克隆水滴伪康纤虫(Pseudocohnilembus persalinus)的钙调素依赖蛋白激酶基因,分析其蛋白结构、功能,并基于该蛋白的氨基酸序列构建相关纤毛虫的系统进化树,以阐明水滴伪康纤虫CaMK的进化特征及与相关纤毛虫的系统发育关系。【方法】参考水滴伪康纤虫RNA-seq测序结果,利用cDNA末端快速克隆(RACE)技术获得水滴伪康纤虫钙调素依赖蛋白激酶基因的全长cDNA序列,将其命名为PpCaMK1;利用生物信息学方法对该基因的序列特征、基因结构进行分析,预测该基因编码蛋白的理化性质、结构域及蛋白结构等,并构建其与相关纤毛虫CaMK的系统发育树。【结果】PpCaMK1基因包含有9个外显子和8个内含子,其cDNA全长为1737 bp(GenBank序列号:PQ278249),其5′-UTR长度为56 bp,3′-UTR长度为307 bp,ORF总长度为1374 bp,编码457个氨基酸残基。PpCaMK1总亲水性平均值为-0.802,不稳定指数为54.78,为亲水性不稳定蛋白。亚细胞定位预测分析表明PpCaMK1存在于细胞质-细胞核中。该蛋白无跨膜结构域,无信号肽。蛋白质的二级结构分析结果显示该蛋白含有的无规卷曲占比达50.33%。构建的系统发育树显示,水滴伪康纤虫PpCaMK1与寡膜纲纤毛虫同源蛋白具有较近的亲缘关系。【结论】本研究克隆了水滴伪康纤虫PpCaMK1基因的全长cDNA,阐明该基因的结构,并初步分析PpCaMK1蛋白的基本理化性质及结构特征,结果可作为进一步研究PpCaMK1功能的基础,并为深入探究水滴伪康纤虫钙调素依赖蛋白激酶基因家族提供参考。【Objective】Gene associated with the calmodulin-dependent protein kinases(CaMKs)from Pseudocohnilembus persalinus was cloned to study its protein structure and function as well as genetic relation and evolutionary characteristics for the development of treatment for the parasitic scuticociliatosis in aquaculture species.【Method】Based on the RNA-seq of P.persalinus,the full-length cDNA sequence of PpCaMK1 was obtained using the Rapid Amplification of cDNA Ends(RACE)technique.Bioinformatic methods were employed to analyze the features,structure,physicochemical properties,domains and phylogenetic relationships with other ciliates of the gene.【Result】PpCaMK1 contained 9 exons and 8 introns.The full cDNA length was 1737 bp(GenBank accession number:PQ278249)including a 5'-UTR of 56 bp,a 3'-UTR of 307 bp,and an ORF of 1374 bp and encoded 457 amino acids.It was a hydrophilic and unstable protein with an average overall hydrophilicity of-0.802 and an instability index of 54.78.Distributed within the cytoplasm and nucleus,PpCaMK1 notably lacked the transmembrane domains and signal peptides and had a secondary structure composed of a significant proportion(i.e.,50.33%)of random coils.P.persalinus closely related to other ciliates in the class Oligohymenophorea.【Conclusion】The full-length cDNA of PpCaMK1 was successfully cloned and the basic biological and physicochemical characteristics studied paving the way for further research on the development of an effective treatment of the devastating disease on marine lives in aquacultural environment caused by the parasite P.persalinus.

关 键 词：水滴伪康纤虫 CAMK 基因克隆 生物信息学 

分 类 号：Q956[生物学—动物学]