sod基因缺失对单增李斯特菌10403S抗氧化应激作用的影响  

Effect of sod Gene Deletion on the Anti-oxidative Stress in Listeria monocytogenes 10403S

作　　者：洪嘉俊 肖金华 张政 杨雨婷 方春 韩旭 HONG Jiajun;XIAO Jinhua;ZHANG Zheng;YANG Yuting;FANG Chun;HAN Xu(College of Animal Science and Technology,Yangtze University,Jingzhou 434025,China;Jiangxi Biotech Vocational College,Nanchang 330200,China)

机构地区：[1]长江大学动物科学技术学院,荆州434025 [2]江西生物科技职业学院,南昌330200

出　　处：《中国畜牧兽医》2025年第4期1776-1783,共8页China Animal Husbandry & Veterinary Medicine

基　　金：湖北省教育厅科学技术研究项目(Q20221302);江西省教育厅科学技术研究项目(GJJ2207208)。

摘　　要：【目的】通过构建sod基因缺失株,阐明超氧化物歧化酶编码基因sod对单增李斯特菌10403S抗氧化应激能力的影响。【方法】本研究采用同源重组方法构建sod基因缺失株。通过生长试验分析亲本株10403S、缺失株10403SΔsod、回补株10403S CΔsod的生长能力。通过氧化应激试验测定sod基因对单增李斯特菌氧化应激能力的影响。通过Western blotting测定氧化应激条件对SOD蛋白表达水平的影响。【结果】PCR结果显示,10403SΔsod和10403S CΔsod目的片段大小分别为1 468和1 875 bp,符合预期,成功构建sod基因缺失株10403SΔsod和回补株10403S CΔsod。生长试验结果显示,sod基因缺失不影响10403S的生长能力。抗氧化应激试验结果显示,在20 mmol/L H_(2)O_(2)条件下,sod基因缺失极显著降低了10403S的抗氧化应激能力(P<0.01)。Western blotting结果显示,在氧化应激条件下单增李斯特菌SOD蛋白的表达水平极显著上调(P<0.01)。【结论】sod基因缺失不影响单增李斯特菌10403S正常生长,但会导致氧化应激能力下降,且在20 mmol/L H_(2)O_(2)条件下10403S中SOD蛋白表达水平极显著升高。【Objective】In this study,the sod gene deletion strain was constructed to elucidate the effect of sod gene on the anti-oxidative stress ability of Listeria monocytogenes 10403S.【Method】The sod gene deletion strain was constructed by homologous recombination method.The growth ability of parent strain 10403S,deletion strain 10403SΔsod and complemented strain 10403S CΔsod were analyzed by growth test.The effect of sod gene on oxidative stress of Listeria monocytogenes was determined by oxidative stress test.The effect of oxidative stress on SOD protein expression was determined by Western blotting.【Result】The results of PCR showed that the target fragments of 10403SΔsod and 10403SCΔsod were 1468 and 1875 bp,respectively,deletion strain 10403SΔsod and complemented strain 10403S CΔsod were successfully constructed.The results of growth test showed that sod gene deletion did not affect the growth ability of 10403S.The results of anti-oxidative stress test showed that sod gene deletion reduced the anti-oxidative stress ability of 10403S under the condition of 20 mmol/L H_(2)O_(2)(P<0.01).The results of Western blotting showed that the expression level of SOD protein of Listeria monocytogenes was significantly up-regulated under oxidative stress(P<0.01).【Conclusion】The deletion of sod gene did not affect the normal growth of Listeria monocytogenes 10403S,but it could lead to a decrease in oxidative stress ability,and the expression level of SOD protein in 10403S was significantly increased under the condition of 20 mmol/L H_(2)O_(2).

关 键 词：单增李斯特菌 超氧化物歧化酶(SOD) 氧化应激 多克隆抗体 

分 类 号：S852.61[农业科学—基础兽医学] Q93[农业科学—兽医学]