Construction of a mutant strain of Nocardia farcinica with mce4A gene deletion and analysis of the role of mce4A gene
10.3760/cma.j.issn.0254-5101.2019.01.007
- VernacularTitle:鼻疽诺卡菌mce4A基因缺失株的构建及其相关功能研究
- Author:
Heqiao LI
1
;
Zhenjun LI
;
Qi XIAO
;
Han SONG
;
Lina SUN
;
Xingzhao JI
;
Yongliang LOU
Author Information
1. 温州医科大学检验医学院、生命科学学院 325035
- Keywords:
Nocardia farcinica;
Gene knockout;
mce4A gene;
Bacterial survival;
Adhesion and invasion
- From:Chinese Journal of Microbiology and Immunology
2019;39(1):35-41
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a mutant strain of Nocardia farcinica ( N. farcinica ) IFM10152 with mammalian cell entry 4A gene (mce4A) deletion and to analyze the function of that gene dur-ing infection. -ethods The mutant strain of N. farcinica was constructed through in-frame deletion without antibiotic labeling and verified by PCR and sequencing analysis. To analyze the function of mce4A gene in the interaction between N. farcinica and host cells, in vitro growth experiment, macrophage killing experi-ment using THP-1 ( a human leukemia mononuclear cell line) as the model and adhesion and invasion exper-iments using HeLa cells ( cervical cancer epithelial cells) were carried out. Results The mutant strain with mce4A gene deletion was successfully constructed and named △mce4A. No significant difference in growth rate was observed between the mutant and the wild-type strains. After knocking out the mce4A gene, the ability of N. farcinica to resist macrophage killing was obviously weakened as well as its ability to adhere and invade. Conclusions The mutant strain of N. farcinica with mce4A gene deletion was successfully construc-ted. The mce4A gene might play an important role in the adhesion and invasion of N. farcinica to host cells and its survival in macrophages.
