PCV2 strain with GAS-like motif mutation as well as preparation method and application of PCV2 strain

A technology of motifs and mutation sites is applied in the field of construction of porcine circovirus type 2 mutant strains, which can solve the problems of difficult replication ability weakened PCV2 mutant strains and unresearched virus replication.

Active Publication Date: 2021-04-30
NORTHWEST A & F UNIV
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  • Application Information

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Problems solved by technology

[0004] But for PCV2, whether the host cell protein STAT5 binds to the virus protein or DNA, and what effect it has on virus replication has not been studied
At the same time, it is still difficult to effectively obtain mutant PCV2 strains with significantly weakened replication ability through genetic engineering.

Method used

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  • PCV2 strain with GAS-like motif mutation as well as preparation method and application of PCV2 strain
  • PCV2 strain with GAS-like motif mutation as well as preparation method and application of PCV2 strain
  • PCV2 strain with GAS-like motif mutation as well as preparation method and application of PCV2 strain

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Embodiment Construction

[0042] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. The examples are only used to explain the present invention, rather than to limit the protection scope of the invention.

[0043] The present invention first obtains porcine STAT5 protein (pSTAT5) by molecular cloning, and confirms that pSTAT5 interacts with capsid protein Cap of wild-type PCV2 virus through co-immunoprecipitation test, and further finds that the interaction between pSTAT5 and Cap is indirect through GST-pull down identification. of. The key region of PCV2 virus DNA binding to pSTAT5 was analyzed by sequence alignment, site-directed mutagenesis primers were designed according to the binding site, and PCV2 virus DNA with binding site mutation was amplified by overlapping PCR using wild-type PCV2 DNA as a template. The mutated PCV2 viral DNA is circularized and transfected into PK-15 cells. After culturing for a certain period of...

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Abstract

The invention discloses a PCV2 strain with GAS-like motif mutation as well as a preparation method and application of the PCV2 strain. A GAS-like motif in PCV2DNA is mutated, so that the mutated PCV2DNA cannot be combined with STAT5 protein in virus replication, and a mutant strain of which the replication capacity is far lower than that of a wild PCV2 strain is obtained through virus rescue. The mutant strain can be used for researching a PCV2 replication mechanism and attenuated live vaccines, so that a new thought is provided for prevention and control of PCV2.

Description

technical field [0001] The invention relates to the construction of a porcine circovirus type 2 (PCV2) mutant strain, in particular to a PCV2 virus with a GAS-like motif mutation. Background technique [0002] PCV2 infection causes the body to produce immunosuppression, leading to PCV2 and other pathogens, such as porcine reproductive disorder and respiratory syndrome virus (PRRSV), swine fever virus (CSFV), porcine pseudorabies virus (PRV) and porcine parvovirus (PPV), etc. Mixed infection also reduces the protection of infected pigs to vaccines, which seriously threatens the development of pig industry. Although PCV2 can replicate in PK-15 cells, the mechanism of PCV2 replication is not clear. The study of PCV2 DNA replication mechanism has become one of the key links in the current study of PCV2 replication mechanism. [0003] Signal transducer and activator of transcription 5 (STAT5) belongs to the signal transducer and activator of transcription protein family and is ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/34C12N5/071C12R1/93
CPCC12N7/00C12N5/0686C07K14/005C12N2750/10021C12N2750/10022C12N2750/10052C12N2750/10062Y02A50/30
Inventor 黄勇童德文韩聪杜谦朱磊
Owner NORTHWEST A & F UNIV
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