中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 半滑舌鳎源哈维氏弧菌分泌蛋白HutZ的功能鉴定及免疫原性 水产学报 2022,46 (1) 116-125
Position: Home > Articles > Function and immunogenicity of the secretory protein HutZ of Vibrio harveyi from Cynoglossus semilaevis Journal of Fisheries of China 2022,46 (1) 116-125

半滑舌鳎源哈维氏弧菌分泌蛋白HutZ的功能鉴定及免疫原性

作  者：
尚玉曼;王恒;刘静茹;袁增智;孙金生
单  位：
天津师范大学生命科学学院;天津师范大学天津市动植物抗性重点实验室
关键词：
半滑舌鳎;哈维氏弧菌;HutZ蛋白;免疫
摘  要：
哈维氏弧菌为我国海水鱼类养殖过程中的常见病原,为了进一步探究哈维氏弧菌的致病机制,本实验分别从前期筛选到的哈维氏弧菌强毒株H2LB1和弱毒株H1SAI3中提取胞外产物(ECPs),并通过蛋白质组学进行差异蛋白鉴定。从中选取了哈维氏弧菌强毒株特异性蛋白HutZ为研究对象,对其功能和免疫原性开展深入研究。首先利用qRT-PCR检测了哈维氏弧菌在不同铁源培养基培养条件下的HutZ基因表达量差异,进一步将克隆HutZ基因连接到原核表达载体上,并在大肠杆菌中成功表达出重组HutZ蛋白。将纯化的HutZ蛋白与血红素共孵育,检测发现该蛋白能够结合血红素并改变其光吸收峰。将HutZ蛋白乳化后免疫半滑舌鳎2次,以间接血凝法检测免疫后1—7周的血清效价,同时在免疫4周后进行攻毒检测其免疫保护率。研究表明,HutZ蛋白免疫后的半滑舌鳎抗体效价最高可达1∶128,攻毒免疫保护率达73.3%,说明HutZ蛋白可作为开发预防半滑舌鳎感染哈维氏弧菌的亚单位疫苗的有效候选抗原。
译  名：
Function and immunogenicity of the secretory protein HutZ of Vibrio harveyi from Cynoglossus semilaevis
作  者：
SHANG Yuman;WANG Heng;LIU Jingru;YUAN Zengzhi;SUN Jinsheng;College of Life Sciences, Tianjin Normal University;Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University;
关键词：
Cynoglossus semilaevis;;Vibrio harveyi;;HutZ protein;;immunity
摘  要：
Vibrio harveyi is a pathogen common in the marine fish culture of China. To explore the pathogenic mechanism of V. harveyi and develop effective methods for prevention and control V. harveyi infection, we extracted extracellular products(ECPs) from the high virulent strain H2 LB1 and the low virulent strain H1 SAI3 of V.harveyi. The proteins of ECPs were analyzed and identified by using the label-free proteomics method. According to the result of protein identification, we selected the high virulent strain-specific protein HutZ for further analysis.Firstly, qRT-PCR technology was used to detect the expression difference of of V. harveyi gene HutZ under culture conditions with different iron sources. Secondly,the gene HutZ was cloned to the expression vector,and recombinant HutZ protein was successfully expressed and purified. Then the purified recombinant HutZ protein was incubated with hemin, and the spectrum was scanned at 300-700 nm. Finally, Cynoglossus semilaevis were immunized with purified recombinant HutZ protein twice. After immunization, the serum titers of 1-7 weeks were detected by the indirect hemagglutination method, and the challenge experiment was performed after 4 weeks of immunization. The results showed that the expression of gene HutZ is related to the iron source; HutZ protein affected the absorbance of hemin; the antibody titer of C. semilaevis against HutZ protein reached up to 1∶128,and the immune protection rate was as high as 73.3%. These results indicated that HutZ protein is a vital factor for the iron uptake of V. harveyi and potential candidate antigen for developing effective subunit vaccines to prevent the V. harveyi infection.

相似文章

导出引用
计量
文章访问数: 7
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46