AHFGDS (aeromonas hydrophila five-gene deletion strain) attenuated bacteria without antibiotic markers and application

• Main Authors: Lin Luo, Zhou Yang, Wang Xiaohong, Wei Shun, Li Zhi, Li Jinquan, Yu Wei
• Format: Patent
• Language: Chinese; English
• Published: 24.08.2016
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS; SPIRITS; VINEGAR; WINE

The invention discloses AHFGDS (aeromonas hydrophila five-gene deletion strain) attenuated bacteria without antibiotic markers and an application. Based on high-virulent strain ZYAH72, five virulence factors including aerA, alt, ahp, ast and hly are deleted with a genetic engineering method and a marker-free screening strategy, an AHFGDS with five genes deleted is obtained, and the preservation number of the strain is CCTCC (China Center for Type Culture Collection) No:M2015798. Compared with wild bacteria, the strain can still be subjected to a strong reaction with positive serum even if the pathogenicity is remarkably weakened. Experiments prove that the strain serving as an attenuated live vaccine can effectively protect susceptible aquatic animals against infection of wild AHFGDSs. 本发明公开了种无抗生素标记的嗜水气单胞菌减毒菌及应用。基于强毒力菌株ZYAH72，通过基因工程方法和无标记筛选策略缺失了该菌株中的，，，和五个毒力因子，获得了五基因缺失菌株AHFGDS，该菌株的保藏编号为CCTCC NO:M2015798。申请人发现该菌株与野生菌相比，其致病性显著减弱，但依然能与阳性血清发生强烈反应。经实验证明，将该菌株作为种减毒活疫苗，可以有效保护易感水产类动物抵抗野生嗜水气单胞菌的感染。