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Riemerella anatipestifer gene deletion strain and application thereof

A technology for the deletion of bacteria and genes, which is applied in the fields of immunology and animal molecular biology, and can solve problems such as no products coming out

Inactive Publication Date: 2015-11-11
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, scholars at home and abroad have begun to study R. anatipestifer as a live attenuated vaccine, but so far no relatively mature product has come out.

Method used

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  • Riemerella anatipestifer gene deletion strain and application thereof
  • Riemerella anatipestifer gene deletion strain and application thereof
  • Riemerella anatipestifer gene deletion strain and application thereof

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Embodiment Construction

[0032] The present invention will be described in detail below in conjunction with specific embodiments.

[0033] 1. Cloning of left and right arms of RA-YM06537 / 06542 gene

[0034] According to the gene sequence of RA serotype 1 YM strain (RA-YM) 06537 / 06542 screened by Chen Fenfen (2008), two pairs of primers were designed using Premier 5.0 (all primers were synthesized by Beijing Qingke Xinye Biotechnology Co., Ltd.) and amplified It can be used for the left and right arms of OverlapPCR. A KpnI restriction site was added to the upstream primer L15'-end of Leftarm, and a SacI restriction site was introduced at the downstream primer R25'-end of Rightarm. The primer sequences are as follows:

[0035] L1: 5'-C GGTACC TTTTAACTATTGTATTTTATTTAC-3' (KpnI)

[0036] L2: 5'-CTTTTAAAACTACTGTCCAGTAGATTGATAAGC-3'

[0037] Primers L1 and L2 amplify the upstream homology arm 454bp

[0038] R1: 5'-GTTTCGTTCCACTGTTTTGAAGTTGTTTT-3'

[0039] R2: 5'-TGAGCTCGGAAGGAAGCGTCTTGCAATATT-3'(Sac...

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Abstract

The invention discloses a riemerella anatipestifer gene deletion strain having deletion of a double-component regulator control system 06537 / o6542 gene, and an application of the riemerella anatipestifer gene deletion strain, belonging to the technical field of preparation of animinal genetic engineering vaccines. The riemerella anatipestifer gene deletion strain is preserved in the China Center for Type Culture Collection with the preservation number of CCTCC M 2015306; due to the deletion of the 798bp of the double-component regulator control system 06537 / o6542 gene, the riemerella anatipestifer gene deletion strain has obviously lowered virulence, and the lost capacity of liquefying gelatin; the sequence of the deleted gene is shown as SEQ ID NO:2. After the riemerella anatipestifer gene deletion strain is inoculated to ducklings, the ducklings can have the protective immunity reaction resisting the serum 1 type riemerella anatipestifer virulent strain, so that the infection from the riemerella anatipestife is prevented.

Description

technical field [0001] The invention belongs to the technical field of animal bacterial genetic engineering, and relates to related fields such as animal molecular biology and immunology. Specifically, it relates to a 06537 / 06542 gene deletion strain of the two-component regulatory system of R. anatipestifer and its application. Background technique [0002] Riemerella anatipestifer infection is an acute, contact, septic infectious disease of ducklings caused by Riemerella anatipestifer (RA), due to the high incidence, high mortality and growth retardation of the disease , has become a relatively serious problem in the process of large-scale breeding of meat ducks in my country. At present, 21 serotypes have been reported abroad, and 4 new serotypes have been discovered in China. Currently, only serotype 1 has been found among the RA strains isolated and identified from our province. At present, little is known about the virulence-related genes of R. anatipestifer. The vir...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/01
Inventor 李自力卢媞周祖涛张硕王颖殷学焕毕丁仁刘梅胡思顺金卉石德时肖运才王喜亮许青荣
Owner HUAZHONG AGRI UNIV