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拟穴青蟹Cactus基因的cDNA克隆、序列及生物学功能
引用本文:胡蕾,邓恒为,李晶晶,刘姗姗,何建国,李海云,翁少萍.拟穴青蟹Cactus基因的cDNA克隆、序列及生物学功能[J].水产学报,2020,44(1):21-32.
作者姓名:胡蕾  邓恒为  李晶晶  刘姗姗  何建国  李海云  翁少萍
作者单位:华南农业大学动物科学学院,广东广州 510642;中山大学生命科学学院,有害生物控制与资源利用国家重点实验室,广东广州 510275;中山大学生命科学学院,有害生物控制与资源利用国家重点实验室,广东广州 510275;华南农业大学动物科学学院,广东广州 510642
基金项目:国家自然科学基金(31672677);现代农业产业技术体系专项(CARS-48)
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,实验采用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2035 bp,开放阅读框(ORF)1311 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与甲壳动物聚为一支,与无脊椎动物聚为一大支。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,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。本实验成功扩增了SpCactus基因全长,并进行了生物信息学分析、理化性质预测,初步探讨了其生物学功能,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。

关 键 词:拟穴青蟹  Cactus  基因克隆  组织分布  表达分析
收稿时间:2018/12/28 0:00:00
修稿时间:2019/3/27 0:00:00

Cloning, expression and biological functional characteristics of Cactus from Scylla paramamosain
HU Lei,DENG Hengwei,LI Jingjing,LIU Shanshan,HE Jianguo,LI Haiyun and WENG Shaoping.Cloning, expression and biological functional characteristics of Cactus from Scylla paramamosain[J].Journal of Fisheries of China,2020,44(1):21-32.
Authors:HU Lei  DENG Hengwei  LI Jingjing  LIU Shanshan  HE Jianguo  LI Haiyun and WENG Shaoping
Institution:College of Animal Science, South China Agriculture University, Guangzhou 510642, China;State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China,State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China,State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China,State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China,State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China,College of Animal Science, South China Agriculture University, Guangzhou 510642, China and State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China
Abstract: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)of228 bp,an open reading frame of 1311 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.By 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.
Keywords:Scylla paramamosain  Cactus  gene cloning  tissue distribution  expression analysis
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