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Hayati, M., Hosseinzadeh, S., Tabatabaei, M., Hosseini, S. (2018). Cloning and expression of Listeria monocytogenes hly in Lactococcus lactis. , 31(4), 63-72. doi: 10.22092/vj.2018.110836.1318
M. Hayati; S. Hosseinzadeh; M. Tabatabaei; S.M.H. Hosseini. "Cloning and expression of Listeria monocytogenes hly in Lactococcus lactis". , 31, 4, 2018, 63-72. doi: 10.22092/vj.2018.110836.1318
Hayati, M., Hosseinzadeh, S., Tabatabaei, M., Hosseini, S. (2018). 'Cloning and expression of Listeria monocytogenes hly in Lactococcus lactis', , 31(4), pp. 63-72. doi: 10.22092/vj.2018.110836.1318
Hayati, M., Hosseinzadeh, S., Tabatabaei, M., Hosseini, S. Cloning and expression of Listeria monocytogenes hly in Lactococcus lactis. , 2018; 31(4): 63-72. doi: 10.22092/vj.2018.110836.1318

Cloning and expression of Listeria monocytogenes hly in Lactococcus lactis

تحقیقات دامپزشکی و فرآورده‌های بیولوژیک
Article 8, Volume 31, Issue 4, December 2018, Pages 63-72    PDF (1.39 M)
Document Type: Full Research Paper
DOI: 10.22092/vj.2018.110836.1318
Authors
M. Hayati1; S. Hosseinzadeh* 2; M. Tabatabaei1; S.M.H. Hosseini3
1Department of Pathobiology ,School of Veterinary Medicine. Shiraz University, Shiraz, Iran.
2Department of Food Hyggiene and Quality Control , School of Veterinary Medicine. Shiraz University, Shiraz , Iran.
3Immunology department, Razi Vaccine and Serum Research Institute, Shiraz Branch,Agricultural Research ,Education and Extension Organization (AREEO), Shiraz,Iran.
Abstract
The hly gene is one of the most important virulence factors of Listeria monocytogenes encoding listeriolysin O (LLO). LLO is a potent immunogenic protein eliciting both cellular and humoral immunity and has manyother functions besides hemolytic activity. This protein mediates the bacterial escape to the cytoplasm by disruption of phagosomal membrane leading to intracellular survival and proliferation of L. monocytogenes. On the other hand, Lactococcus lactis is a lactic acid bacterium which has many applications in expression of recombinant proteins. Our aim in this study was expression of recombinant LLO in L. lactis strain. Initially, the pTG19-T plasmid which is a T/A cloning vectorwas employed and then pNZ8110 was used as a Lactococcal secretional expression plasmid to clone L. monocytogenese hly gene by restriction enzymes cutting and ligation method. After cloning and propagation in E. coli MC1061 intermediate host, it was electro-transformed into L. lactis. Expression of LLO was confirmed by SDS-PAGE and western blot.
Keywords
Listeriolysin; Listeria monocytogenese; Lactococcus lactis; Cloning
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