Cloning and Expression of Listeria monocytogenes Listeriolysin O in Lactobacillusplantarum

International Journal of Enteric Pathogens

article

Masoumeh Hayati¹  Saeid Hosseinzadeh²  Seyed Mohammad Tabatabaee¹  Seyed Mohammad Hossein Hosseini³  Abdollah Derakhshandeh¹ 

[1] Department of Pathobiology, School of Veterinary Medicine, Shiraz University;Department of Food Hygiene and Quality Control, School of Veterinary Medicine, Shiraz University;Immunology Department, Razi Vaccine and Serum Research Institute, Shiraz Branch, Agricultural Research

关键词: Listeriolysin O;    Cloning;    Lactobacillus plantarum;   

DOI  :  10.15171/ijep.2017.27

学科分类：农业科学（综合）

来源: Alborz University of Medical Sciences

【 摘 要 】

Background: The protein listeriolysin O (LLO) encoded by hly gene, is one of the most important virulence factors of Listeria monocytogenes, responsible for phagosomal membrane disruption and bacterial escape to the cytoplasm, stimulation of CD8+ T cells and Th1 response. Recently pathobiotechnological vaccination using probiotic bacteria have been proposed. One of these strategies is expression of LLO in non-pathogenic bacteria such as lactic acid bacteria as delivery strains. Objective: In the current study, we aimed to clone hly gene in a Lactobacillus species via pNZ8110, an inducible expression vector which is specific for Lactococcus species. Materials and Methods: Hly gene was amplified by polymerase chain reaction (PCR) and inserted into pNZ8110 by restriction enzymes cutting and ligation method. After transformation and propagation in Escherichia coli MC1061 intermediate host, it was successfully electrotransformed into Lactobacillus plantarum. Results: Gel electrophoresis of colony PCR, extracted plasmids and restriction analysis along with sequencing confirmed the transformation. After induction with supernatant of nisin producer, strain Lactococcus lactis NZ9700, expression of LLO was confirmed by SDS-PAGE and western blot. Conclusion: Here, we employed a nonpathogenic probiotic strain, L. plantarum for the first time to express hly gene of L. monocytogenes in order to propose a new vaccine candidate.

【 授权许可】

CC BY-NC