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| 中国明对虾(Fenneropenaeus chinensis)丝氨酸蛋白酶同源物基因(Fc-SPH)的重组表达及活性分析 |
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杨 燚1,2, 刘逸尘1,2, 张亦陈1,2, 孙 妍3, 耿绪云3, 孙金生1,2,3
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1.天津师范大学生命科学学院;2.天津市细胞遗传与分子调控重点实验室;3.天津市水产养殖病害防治中心
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| 摘要: |
| 以实验室前期克隆到的中国明对虾丝氨酸蛋白酶同源物基因(Fc-SPH)(GenBank 登录号: DQ318859)为基础, 利用原核表达系统对Fc-SPH基因成熟肽区域进行了重组表达和纯化复性分析, 并对获得的重组目的蛋白开展了抑菌活性及微生物清除功能研究。结果表明: 在体外成功获得了大量有活性的对虾Fc-SPH蛋白(rFc-SPH), 活性研究显示rFc-SPH 对大多数受试菌种都有明显的抑制效果, 并可以加速中国明对虾清除体内外源微生物的速度。作者推断Fc-SPH即可以作为一线防御应答效应物直接参与虾类的先天免疫活动而发挥作用, 也可以通过调理作用促进血细胞对病原微生物的吞噬和杀灭。 |
| 关键词: 中国明对虾, 丝氨酸蛋白酶同源物, clip 结构域, 生物活性, 先天免疫 |
| DOI：10.11693/hyhz201204024024 |
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| 基金项目:国家重点基础研究发展计划项目(2012CB114405)；国家自然科学基金项目(30901094 和30901119); 国家科技支撑计划项目(2011BAD13B04 和2011BAD13B07); 天津市自然科学基金项目(10JCZDJC18200 和10JCYBJC09200) |
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| RECOMBINANT EXPRESSION AND BIOACTIVITY ANALYSIS OF A SERINE PROTEINASE HOMOLOGUE GENE (Fc-SPH) FROM THE CHINESE SHRIMP FENNEROPENAEUS CHINENSIS |
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YANG Yi1,2, LIU Yi-Chen1,2, ZHANG Yi-Chen1,2, SUN Yan3, GENG XU-Yun3, UN Jin-Sheng1,2,3
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1.College of Life Science, Tianjin Normal University;2.Tianjin Key Laboratory of Cyto-genetical and Molecular Regulation;3.Tianjin Aquaculture Disease Prevention & Treatment Center
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| Abstract: |
| Like other crustaceans, shrimp have a non-specific innate immunity which is composed of diverse processes and molecules to defend themselves against invading pathogens. Arthropod clip domain serine proteinase homologues (clip-SPHs) have been shown to be involved in various biological functions including innate immunity. A serine proteinase homologue (Fc-SPH) cDNA was cloned from the haemocytes of Fenneropenaeus chinensis. Fc-SPH contains a trypsin-like serine protease domain (Tryp_SPc domain) and a clip domain. The transcript of Fc-SPH is induced in response to white spot syndrome virus (WSSV) infection. Recombinant Fc-SPH (rFc-SPH) were successfully expressed in bacteria and purified for further research of biological functions. It showed in vitro antimicrobial activity against the Bacillus thurigiensis, Bacillus subtilis,Vibrio anguillarum,Escherichia coli, Vibrio harveyi and Saccharomyces cerevisiae. A higher bacterial clearance rate of V. harveyi coated with the rFc-SPH compared with V. harveyi only was shown by in vivo study. It revealed that Fc-SPH act as opsonin and could help shrimp to remove external microorganism. The results suggest that Fc-SPH might be a multifunctional immune molecule in shrimp defense. It can provide us clues to understand the probable role of SPH in innate immunity of shrimp. |
| Key words: Fenneropenaeus chinensis, Serine proteinase homologue (SPH), clip domain, Bioactivity, Innate immunity |