首页 > 过刊浏览>2024年第43卷第1期 >203-209. DOI:10.13300/j.cnki.hnlkxb.2024.01.023
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外膜蛋白OmpA在蛙源米尔伊丽莎白菌致病性中的功能
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作者:
作者单位:

华中农业大学水产学院/湖北省水生动物病害防控工程技术研究中心/ 农业农村部水生动物疫病专业实验室(华中农业大学)，武汉430070

作者简介:

刘芳园，E-mail： liufangyuan@webmail.hzau.edu.cn

通讯作者:

顾泽茂，E-mail： guzemao@mail.hzau.edu.cn

中图分类号:

S941.42

基金项目:

国家自然科学基金项目（32202990）；湖北省农业科技创新中心（2021-620-000-001-33）；湖北省自然科学基金项目（2021CFB128）；中国博士后科学基金项目（2021M701351）


Function of outer membrane protein A in pathogenicity of Elizabethkingia miricola from frog
Author:
Affiliation:

College of Fisheries， Huazhong Agricultural University/Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention/ National Aquatic Animal Diseases Para-Reference Laboratory (HZAU)， Wuhan 430070， China

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    摘要:

    为探究外膜蛋白A（outer membrane protein A，OmpA）对米尔伊丽莎白菌致病作用的影响，以蛙源米尔伊丽莎白菌FL160902为研究对象，通过同源重组法构建OmpA缺失株△ompA，比较缺失株和野生株的生长特性、生物膜形成能力、抗血清杀伤能力、对细胞的黏附能力以及对蛙的致病性差异。结果显示：△ompA的生长能力和抗血清杀伤能力与野生株无显著差异；但与野生株相比，△ompA的生物膜形成能力增加了66%，△ompA对bEnd.3细胞的黏附能力降低了61%；黑斑蛙感染试验显示，△ompA在黑斑蛙血液、脾和脑组织中的载菌量分别为（3.15×108±0.09×108）、（2.11×108±0.07×108）和（6.61×108±0.16×108） copies/g，均显著低于野生株，且△ompA对黑斑蛙的致死率为37%，显著低于野生株的致死率（75%）。上述结果表明，ompA基因缺失不改变米尔伊丽莎白菌的抗血清杀伤能力，但增加了菌株的生物膜形成能力，减弱了菌株的黏附能力，从而降低了该菌对蛙的致病性。

    Abstract:

    To study the role of outer membrane protein A （OmpA） in the pathogenicity of Elizabethkingia miricola， the ompA gene mutant strain （△ompA） was constructed via homologous recombination using E. miricola FL160902 as the wild type strain. Subsequently， the growth characteristics， biofilm formation， serum survival ability， adhesion ability， bacterial colonization of the wild strain and △ompA， as well as their pathogenicity to frogs were analyzed. The results showed that there was no significant difference in growth characteristics and survive ability in frog serum between △ompA and wild strain. However， compared with the wild strain， the biofilm formation capacity of △ompA increased by 66%， and the adhesion capacity of △ompA to bEnd.3 cells decreased by 61%. The results of animal experiment showed that the average bacterial load of △ompA in the blood， spleen and brain of infected frogs were （3.15×108±0.09×108）， （2.11×108±0.07×108） and （6.61×108±0.16×108） copies/g， respectively， which were significantly lower than that of the wild strain infected frogs （P<0.05）， and the lethality rate of frogs infected with △ompA was 37%， which was significantly lower than that of the wild strain （75%）. The results indicated that ompA deletion did not change the antiserum killing ability of E. miricola， but increased the biofilm forming ability of E. miricola， reduced its adhesion ability and decreased its pathogenicity to frogs.

    表 1 引物信息Table 1 Primers information
    图1 缺失株△ompA的PCR鉴定Fig.1 Identification of the ompA mutant strain by PCR
    图2 野生株和缺失株△ompA的生长曲线Fig.2 Growth curve of the wild-type strain and mutant strain ΔompA
    图3 野生株和缺失株△ompA的生物膜形成能力Fig.3 Biofilm formation capacities of the wild-type strain and mutant strain △ompA
    图4 野生株和缺失株△ompA在50%血清中的存活能力Fig.4 Survival rate of the wild-type strain and mutant strain △ompA in 50% serum
    图5 野生株和缺失株△ompA黏附bEnd.3细胞能力Fig.5 Adherence abilities of the wild-type strain and mutant strain △ompA
    图6 感染后黑斑蛙血液（A）、脾（B）和脑组织（C）中载菌量Fig.6 Bacterial loads in blood （A）， spleen （B） and brain （C） of infected frogs
    图7 黑斑蛙感染野生株和缺失株△ompA后的存活率Fig.7 Survival rate of frogs infected with the wild-type strain and mutant strain △ompA
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刘芳园,胡瑞雪,余芳,侯家昊,于子润,顾泽茂.外膜蛋白OmpA在蛙源米尔伊丽莎白菌致病性中的功能[J].华中农业大学学报,2024,43(1):203-209

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  • 收稿日期:2022-11-22
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