文章摘要
拟穴青蟹Cactus基因的cDNA克隆、序列及生物学功能
Cloning, Expression and Biology Functional Characteristic of Cactus form Scylla paramamosain
投稿时间:2018-12-28  修订日期:2019-03-27
DOI:
中文关键词: 拟穴青蟹  Cactus  基因克隆  组织分布  表达分析
英文关键词: Scylla paramamosain  Cactus  gene cloning  tissue distribution  expression analysis
基金项目:国家自然科学基金
作者单位E-mail
胡蕾 动物科学学院 369359469@qq.com 
邓恒为 中山大学生命科学学院  
李晶晶 中山大学生命科学学院  
刘姗姗 中山大学生命科学学院  
何建国 中山大学生命科学学院  
李海云 动物科学学院  
翁少萍 中山大学生命科学学院 lsswsp@mail.sysu.edu.cn 
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中文摘要:
      为获取拟穴青蟹Cactus基因cDNA全长、分析基本生物学信息,并初步探索其在病原物刺激下的免疫反应。本文采用 RACE 技术获得了拟穴青蟹 Cactus(SpCactus)基因的 cDNA 全长序列,其 cDNA 全长为1 855 bp,开放阅读框(ORF) 1 311 bp,编码436个氨基酸,分子量为46.01 ku。对蛋白理化性质进行预测发现,SpCactus为亲水性蛋白,等电点pI为 4.91。经预测,SpCactus与其他物种的IκB蛋白具有相似的功能结构域。同源性比对结果显示,SpCactus与凡纳滨对虾和中国明对虾的Cactus蛋白同源性均高达62%,相似度为73%。系统进化树分析显示SpCactus与甲壳动物聚为一类,与无脊椎动物聚为一大类。实时荧光定量 PCR(RT-PCR)检测发现SpCactus基因在拟穴青蟹不同组织中均有表达,肌肉中的表达量最高,其次为心脏、眼柄、血液和鳃组织,肝胰腺中的表达量最低。LPS和金黄色葡萄球菌刺激均能显著诱导SpCactus 基因的表达。本实验成功扩增SpCactus基因全长,并进行生物信息学分析、理化性质预测,初步探讨其生物学功能,为进一步研究其在免疫反应中的生物学功能提供参考依据。
英文摘要:
      In this study, We obtained the full length of the spcactus gene by RACE technology from Scylla paramamosain(SpCactus). And we analyzed the biological information of the SpCactus and its immune response under pathogen stimulation. The full length of SpCactus mRNA contains a 5′ untranslated region (UTR) of 228 bp, an open reading frame of 1 311 bp and a 3′ UTR of 496 bp. The SpCactus protein contains five characteristic ANK homology domains and showed 62% identity (73% similarity), 24% identity (36% similarity), and 9% identity (18% similarity) to the Litopenaeus vannamei Cactus (LvCactus) protein, the Drosophila melanogaster Cactus (DmCactus), and the Homo sapiens IκB protein, respectively. Prediction of the protein physicochemical properties of SpCactus revealed that it is a hydrophilic protein. The physical and chemical properties analysis, SpCactus was a hydrophilic protein. And its isoelectric point (pI) is 4.91. The mRNA of SpCactus showed high expression in the muscle, Eyestalk and Heart, but low expression in the hepatopancreas. Moreover, the expression of SpCactus were significantly upregulated by stimulation with Staphylococcus aureus and lipopolysaccharide. In this study, SpCactus gene was successfully cloned and characterized for the first time, and its bioinformatics analysis, physical and chemical properties were predicted. The biological functions of SpCactus gene were preliminarily explored, providing a basis for further study of its biological functions in immune response.
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