赤子爱胜蚓Calreticulin的cDNA和氨基酸序列分析

目的分析赤子爱胜蚓Calreticulin的cDNA和氨基酸序列。方法从本院构建的赤子爱胜蚓cDNA文库中经测序获得其Calreticulin cDNA序列。采用生物信息学分析工具DNAMAN、NCBI ORF finder、ExPASy Protparam、ExPASY Protscale、NCBI Conserved Domains、SignalP 4.1 Server、TMHMM Server v.2.0、SOSUI、NetPhos 3.1 Server、PSORT、SOPMA、ExPASy COILS、SWISS-MODEL分别对Calreticulin的cDNA序列、氨基酸序列、一级结构基本理化性质、亲/疏水性、保守结构域、信号肽、跨膜区、可溶性、磷酸化修饰、亚细胞定位、二级结构、卷曲螺旋域以及三级结构进行预测及分析。结果赤子爱胜蚓Calreticulin cDNA序列长476 bp,其中包含354 bp的开放阅读框,编码118个氨基酸残基;Calreticulin由19种氨基酸组成,呈酸性,为稳定的可溶性亲水蛋白;具有1段Calreticulin超家族保守结构域;不存在信号肽和跨膜结构;具有12个潜在的磷酸化位点;主要存在于细胞核中;二级结构元件主要由α-螺旋(29.66%)和无规则卷曲(50.85%)组成,具有卷曲螺旋;三级结构预测结果与二级结构一致。结论掌握了赤子爱胜蚓Calreticulin的理化性质,预测了其潜在的特征,并分析了其结构,为后续深入开展其功能及分子作用机制的研究提供了理论依据。

Analysis of cDNA and amino acid sequences of Calreticulin from Eisenia fetida

Objective To analyze the cDNA and amino acid sequences of Calreticulin from Eisenia foetida.Methods The cDNA sequence of Calreticulin was obtained from the Eisenia foetida cDNA library constructed.By using the bioinformatic tools such as DNAMAN,NCBI ORF finder,ExPASy Protparam,ExPASY Protscale,NCBI Conserved Domains,SignalP 4.1 Server,TMHMM Server v.2.0,SOSUI,NetPhos 3.1 Server,PSORT,SOPMA,ExPASy COILS and SWISS-MODEL,the sequences of Calreticulin cDNA and amino acid,physicochemical properties of primary structure,hydrophilicity/hydrophobicity,conservative domain,signal peptide,transmembrane domain,solubility,phosphorylation modification,subcellular localization,secondary structure,coiled helix domain and tertiary structure were predicted and analyzed.Results The Calreticulin cDNA sequence of Eisenia foetida was at a length of 476 bp,which contained a 354 bp open reading frame(ORF)encoding 118 amino acid residues.Calreticulin consisted of 19 kinds of amino acids,which was acidic and stably soluble hydrophilic protein.It had a conserved domain of Calreticulin superfamily while no signal peptide or transmembrane structure,but had 12 potential phosphorylation sites.It was mainly located in the cellnucleus,of which the secondary structure was mainly alpha-helix(29.66%)and random coil(50.85%),with coiled helical domain,while the predicted result of tertiary structure was consistent with that of secondary structure.Conclusion The physicochemical properties of Calreticulin from Eisenia foetida were understood,while the potential characteristics were predicted,and the structure was analyzed,which provided a theoretical basis for further study on the function and molecular mechanism of Calreticulin.