布鲁氏菌ABC转运蛋白相关基因cydDC缺失株的构建及生物学特性分析

doi:10.16431/j.cnki.1671-7236.2025.11.029

Abstract

Abstract: 【Objective】 This study aimed to investigate the biological functions of cydDC gene in Brucella,and reveal its role in stress response and intracellular survival. 【Method】 Using Brucella melitensis 16M as the model organism,cydDC gene deletion strain (16MΔcydDC) was constructed through homologous recombination,and a complemented strain (16M CΔcydDC) was generated by transforming complementation plasmid.By comparison of growth characteristics,stress sensitivity,intracellular survival capacity,and inflammatory cytokine induction among these three strains,the functional role of the cydDC gene was comprehensively evaluated. 【Result】 PCR analysis identified a 4 504 bp amplicon in wild type 16M,while both 16MΔcydDC and 16M CΔcydDC strains showed a 1 088 bp fragment,with the complemented strain additionally exhibiting a 3 819 bp product,confirming successful mutant construction.Although all three strains displayed comparable growth kinetics in vitro,stress susceptibility assays demonstrated significantly enhanced sensitivity of the 16MΔcydDC strain to both acidic (pH 4.0-4.5) and oxidative (1 mmol/L H₂O₂) challenges compared with wild-type and complemented strains (P＜0.05).Intracellular proliferation assays further revealed that at 24 h post-infection, 16MΔcydDC strain showed significantly impaired survival in RAW264.7 macrophages (P＜0.05),concomitant with markedly upregulated expression of proinflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) (P＜0.05). 【Conclusion】 ABC transporter-related gene cydDC contributed to Brucella survival under acidic and oxidative stress,as well as intracellular environments,and reduced the damage caused by such stimuli to the bacteria by regulating cellular inflammatory responses.

Key words: Brucella; cydDC gene; gene deletion; biological characteristics

CLC Number:

S852.61

JIANG Zhiqiang, ZHANG Haobo, LIU Mengda, SUN Xiangxiang, SHAO Weixing, CHANG Lu, QI Fei, SUN Shixiong, YAN Xin, NAN Wenlong, SUN Shufang, FAN Xiaoxu, HUANG Baoxu. Construction and Biological Characterization Analysis of Brucella ABC Transporter-related Gene cydDC Deletion Strain[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5338-5349.

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Construction and Biological Characterization Analysis of Brucella ABC Transporter-related Gene cydDC Deletion Strain

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