Transformation and identification of lipid A modified Coxiella burnetii mutant
WANG Tao
WANG Quan
LUO Shengdong
ZHANG Aimin
LI Wengang
DUAN Xuezhang
Abstract：Objective To explore an experimental laboratory model of lipopolysaccharides modified Coxiella mutant,based on the electroporation technique of post genetic cloning.Methods The shuttle vector was constructed by designing and introducing the chlamydial KDO gene through molecular cloning.After electroporating Coxiella burnetii,clonal purification and expansion were performed using a semi-solid axenic medium.Additionally,the effects of LPC-011 inhibitors on Coxiella were observed using phase-contrast microscopy.Comparative growth curves of the transformed mutant and wild strain were measured under cellular and axenic conditions using qPCR quantitative analysis,and the infectivity of CFUs was detected in Vero cells.Results The shuttle vector pCBGkdtA can stably transform Coxiella.The minimal inhibitory concentration(MIC)of LPC-011 on Coxiella may be greater than 10 μg/mL.Compared with the wild strain,the growth and propagation character of lipid A-modified Coxiella did not chang apparently.Conclusion CBkdtA,lipid A modified Coxiella transformant was successfully constructed,moreover,both its property of reproducing and infecting were identified.The study is provides a new method of further exploring the pathogenic mechanisms of Coxiella lipopolysaccharides in animal models.
Keywords：Coxiella burnetiilipopolysaccharideslipid Aelectro-transformationgene
Publication Date：2024-10-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 455-458,475 )