鳗弧菌vah1缺失菌株构建及致病性评估  

Construction and pathogenicity evaluation of Vibrio anguillarum strain with vah1 deletion

作　　者：胡存洁 车金远 黄旭雄[1,2] 罗土炎 鲍宝龙[1,2] HU Cunjie;CHE Jinyuan;HUANG Xuxiong;LUO Tuyan;BAO Baolong(Key Laboratory of Exploration and Utilization of Aquatic Genetic Resoures,Ministry of Education,Shanghai Ocean University,Shanghai 201306,China;National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China;Institute of Quality Standards and Testing Technology for Agro-Products,Fujian Academy of Agricultural Sciences,Fuzhou 350003,Fujian,China)

机构地区：[1]上海海洋大学水产种质资源发掘与利用教育部重点实验室,上海201306 [2]上海海洋大学水产科学国家级实验教学示范中心,上海201306 [3]福建省农业科学院农业质量标准与检测技术研究所,福建福州350003

出　　处：《上海海洋大学学报》2023年第3期481-490,共10页Journal of Shanghai Ocean University

基　　金：福建省省属公益类科研院所基本科研专项(2020R1022009);福建省区域发展项目(2021N3016)。

摘　　要：通过基因重组技术,敲除鳗弧菌毒力基因vah1,成功构建vah1缺失鳗弧菌菌株Δvah1。Δvah1菌株溶血活性下降89.4%,生物膜形成能力下降16%。Δvah1菌株感染凡纳滨对虾(Litopenaeus vannamei)的半数致死量为(9.77±0.50)×10^(5)cfu/mL,相比鳗弧菌野生菌株的半数致死量(3.16±0.80)×10^(5)cfu/mL提高了2.09倍;相比鳗弧菌野生菌株,Δvah1菌株在凡纳滨对虾(Litopenaeus vannamei)肝胰腺的定殖能力明显下降,对肝胰腺细胞造成溶解和空泡化的破坏程度也相对较小,肝胰腺免疫相关基因CAT、SOD、PO、LZM和ACP的表达水平降低。这些研究结果表明,vah1缺失影响鳗弧菌对凡纳滨对虾的致病能力,Δvah1菌株为后续鳗弧菌减毒疫苗或抗鳗弧菌免疫增强剂的制备奠定了基础。The virulence gene vah1 in Vibrio anguillarum was knocked out by gene recombination,and the strainΔvah1 with vah1 deletion was constructed in this study.The hemolytic activity ofΔvah1 strain decreased by 89.4%and the biofilm forming ability decreased by 16%.The median lethal dose(LD 50)ofΔvah1 strain infected Litopenaeus vannamei was(9.77±0.50)×10^(5) cfu/mL,compared with(3.16±0.80)×10^(5) cfu/mL of the LD 50 of the wild strain of V.anguillarum,it increased by 2.09 times;the colonization ability ofΔvah1 strain in L.vannamei decreased evidently after vah1 deletion.Compared with the wild strain of V.anguillarum,the destruction degree ofΔvah1 strain on hepatopancreas cells was significantly smaller.AfterΔvah1 strain infected L.vannamei,the expression levels of immune related genes CAT,SOD,PO,LZM and ACP in hepatopancreas decreased to some extent.These results showed that the hemolysis and pathogenicity to L.vannamei of the vah1 deficient V.anguillarum strain were reduced in varying degrees,which laid a foundation for the preparation of subsequent attenuated vaccines or specific immune enhancers against V.anguillarum.

关 键 词：鳗弧菌 vah1基因 致病性 凡纳滨对虾 减毒菌株 

分 类 号：S917[农业科学—水产科学]