摘要
甲壳动物的血细胞参与多种免疫反应,能够激活酚氧化酶原,产生抗菌肽或进行血细胞的封装、吞噬等.本研究在克氏原螯虾血细胞中发现一种造血激素astakine,是与脊椎动物细胞因子前动力蛋白prokineticin同源的基因,命名为PcAst,该基因的开放阅读框为357 bp,编码118个氨基酸,具有保守的半胱氨酸残基.半定量PCR结果显示,该基因只在血细胞中表达.实时荧光定量PCR结果显示,在金黄色葡萄球菌和鳗弧菌刺激后,其mRNA表达量大幅上调.白斑综合征病毒刺激后,该基因也呈持续上调表达趋势.为进一步研究其功能,重组表达了PcAst蛋白,并对其蛋白水平表达模式进行了分析.所有结果表明,该基因可能参与鳌虾血细胞的固有免疫应答反应,在抗细菌和抗病毒中起重要作用.
Crustacean hemocytes are involved in various immune responses, including prophenoloxidase activation, antimicrobial peptide production and phagocytosis, tn this study we report a novel gene astakine related to hemocytes differentiation, designated as PcAst, from the red swamp crayfish, Procambarus clarkii. This astakine is homologous with the prokineticin gene of vertebrates, and it contains an ORF of 357 bp, encoding a peptide with 118 amino acids with conservative cysteines. The semi-quantitative PCR results showed that PcAst transcripts were only expressed in hemocytes, and quantitative PCR revealed that PcAst was significantly up-regulated after Vibrio anguillarum, Staphyloccocus aureus or white spot syndrome virus challenge. To further study the function, we made PcAst recombinantly expressed in E. coli and the expression profiles of PcAst in protein level were analyzed. Taken these results together, it was proposed that this PcAst gene might be involved in the immune responses of crayfish against bacteria and virus.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2013年第12期1172-1179,共8页
Chinese Journal of Biochemistry and Molecular Biology
基金
河南省科技厅基础与前沿技术研究计划项目(No.132300410140)
河南省教育厅科学技术重点研究项目(No.12A180018)
河南师范大学博士科研启动课题(No.11133)
国家自然科学基金(No.31301883)~~
关键词
固有免疫
细胞因子
鳌虾
表达模式
细菌
病毒
innate immune
cytokine
crayfish
expression pattern
bacteria
virus
