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金黄色葡萄球菌噬菌体裂解酶Lys239表达纯化及活性分析

李君 孙虎芝 田延军 潘强 任慧英 刘文华

动物医学进展2024,Vol.45Issue(9):27-31,5.
动物医学进展2024,Vol.45Issue(9):27-31,5.

金黄色葡萄球菌噬菌体裂解酶Lys239表达纯化及活性分析

Expression,Purification and Activity Analysis of Lys239 Phage Lyase from Staphylococcus aureus

李君 1孙虎芝 2田延军 2潘强 2任慧英 2刘文华3

作者信息

  • 1. 青岛农业大学动物医学院,山东青岛 266109||青岛诺安百特生物技术有限公司,山东青岛 266000
  • 2. 青岛诺安百特生物技术有限公司,山东青岛 266000
  • 3. 青岛农业大学动物医学院,山东青岛 266109
  • 折叠

摘要

Abstract

In order to obtain phage lyase of Staphylococcus aureus with high lytic activityy,the entire gene sequence of the sequenced phage was compared in the GenBank database,and the lytic enzyme gene se-quence was excavated.After cloning,the protein was expressed in BL21(DE3)and purified.The Galleria mellonella was selected as a model to simulate bacterial infection in vivo to explore its antibacterial effect.The results showed that the lyase synthesis gene was successfully constructed into the vector pET22b,re-sulting in a recombinant plasmid pET22b-lys239.And the recombinant plasmid was transfected into BL21(DE3)receptive cells to induce its expression.After purification and validation,Lys239 with lysis activity at a concentration of 2 μg/μL was obtained.The results of the bacterial infection test in the body of the Galle-ria mellonella showed that the survival rate of the Lys239 treatment group was significantly higher than that of the control group.The hemolymph load test showed a significant difference between the control group and each treatment group(P<0.01),and the higher the concentration of Lys239,the more signifi-cance the inhibitory effect on Staphylococcus aureus.The recombinant lysase Lys239 expressed in this study has good antibacterial effects and can be used as a potential new antibacterial agent.

关键词

金黄色葡萄球菌/噬菌体/裂解酶/大蜡螟幼虫/抑菌作用

Key words

Staphylococcus aureus/phage/lyase/Galleria mellonella/bacteriostatic effect

分类

农业科技

引用本文复制引用

李君,孙虎芝,田延军,潘强,任慧英,刘文华..金黄色葡萄球菌噬菌体裂解酶Lys239表达纯化及活性分析[J].动物医学进展,2024,45(9):27-31,5.

基金项目

山东省产业技术体系驴产业创新团队项目(SDAIT-27) (SDAIT-27)

动物医学进展

OA北大核心CSTPCD

1007-5038

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