Promoting resuscitation of viable but nonculturable cells ofVibrio harveyiby a resuscitation-promoting factor-like protein YeaZ
| Autor: | Mingjun zhao, Zhi Yang, Jixiang Chen, Yanlin Li, Liang Yue, Xiao-Hua Zhang |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Resuscitation Mutant Virulence medicine.disease_cause Applied Microbiology and Biotechnology Viable but nonculturable law.invention Microbiology 03 medical and health sciences Bacterial Proteins law medicine Animals Cloning Molecular Escherichia coli Phylogeny Zebrafish Vibrio Microbial Viability biology Vibrio harveyi Temperature General Medicine Suicide gene biology.organism_classification 030104 developmental biology Recombinant DNA Cytokines Biotechnology |
| Zdroj: | Journal of Applied Microbiology. 122:338-346 |
| ISSN: | 1364-5072 |
| Popis: | Aims To demonstrate the resuscitation promoting activities of recombinant YeaZ from V. harveyi SF-1. Methods and Results The gene of resuscitation-promoting factor YeaZ was cloned from genomic DNA of Vibrio harveyi SF-1. The gene was expressed in Escherichia coli and the expressed protein was purified by Ni2+-affinity chromatography. A yeaZ mutant was constructed by using the suicide plasmid pNQ705 with homologous recombination. Disruption of yeaZ did not affect cell growth significantly in 2216 E broth at 28°C. The wild-type and mutant VBNC cells could be resuscitated by temperature upshift method. Addition the recombinant YeaZ increased the culturable counts from 1.27×104 CFU ml−1 and 1.99×104 CFU ml−1 to 2.88×105 CFU ml−1 and 4.59×105 CFU ml−1 respectively. After the VBNC cells of wild-type and mutant cells were maintained at 4°C for 120 days, no resuscitation was obtained by temperature upshift method, but addition of the recombinant YeaZ promoted the resuscitation of the wild-type and mutant cells, with the culturable cell counts of 1.13×103 CFU ml−1 and 1.44×103 CFU ml−1 respectively. Disruption of yeaZ decreased the virulence of V. harveyi in zebrafish. The LD50 of the yeaZ null mutant was more than 10 fold higher than that of the wild-type cells. Conclusions The recombinant YeaZ could efficiently promote resuscitation of the wild-type and mutant cells of V. harveyi from VBNC to culturable state. The protein also promoted resuscitation of the VBNC wild-type and mutant cells, which were maintained at 4°C for 120 days and not recovered by temperature upshift method. Disruption of yeaZ decreased the virulence of V. harveyi in zebrafish. Significance and Impact of the Study Here we show clear evidence of a resuscitation promoting factor YeaZ of V. harveyi and the roles in resuscitation of the VBNC cells and its pathogenicity. This article is protected by copyright. All rights reserved. |
| Databáze: | OpenAIRE |
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