5 results about "Cosmid" patented technology

The present invention provides a recombinant Marek's disease virus that co-expresses the Newcastle disease virus F gene and the infectious bursal disease virus new variant VP2 gene and its application, belonging to the field of genetically engineered vaccines. The present invention utilizes recombinant cloning technology to insert an expression framework comprising the Newcastle disease virus gene type VII strain F gene and the infectious bursal disease virus new variant VP2 gene into the serotype 1 chicken Marek's disease virus CVI988 strain genome between nucleotides 105561-105563, constructing a recombinant cosmid in which the Newcastle disease virus F gene and the infectious bursal disease virus new variant VP2 gene expression framework are inserted into the chicken Marek's disease virus genome, and the recombinant virus C20103 strain that co-expresses the Newcastle disease virus F gene and the infectious bursal disease virus new variant VP2 gene is rescued by the recombinant Marek's disease virus. The vaccine prepared by the recombinant chicken Marek's disease virus provided by the present invention can simultaneously prevent Newcastle disease, infectious bursal disease and chicken Marek's disease, and has good market application prospects.

This invention belongs to the field of viral genetic engineering technology and discloses a bovine herpesvirus type 1 (BHV-1) with three gene deletions, its construction method, and its applications. Using a Fosmid library, four myxovirus rescue combinations were screened. FosBHV-1-b-ΔTK-mCherry and FosBHV-1-d-ΔgEct / US9-eGFP were obtained through Red / ET and Gateway methods. These were then combined with FosBHV-1-a and FosBHV-1-c to construct an infectious cloning platform for recombinant BHV-1 with the TK / gEct / US9 three-gene deletions, rescuing recombinant viruses carrying tracer genes. This method has high recombination efficiency, and the cloning platform can efficiently construct BHV-1 gene-deleted strains. The immunogenicity of the deleted strains is comparable to that of wild-type viruses, with good safety, providing an efficient technical means for the development of BHV-1 attenuated vaccines and bovine disease live vector vaccines.

The invention provides a bovine herpesvirus type 1 (BHV-1) infectious clone as well as a construction method and application thereof, and belongs to the technical field of virus genetic manipulation. According to the invention, a group of cosmid combinations for constructing the BHV-1 infectious clone is screened based on a Fosmid library, and the cosmid combinations comprise FosBHV-1-a, FosBHV-1-b, FosBHV-1-c and FosBHV-1-d. According to the cosmid combination, BHV-1 can be efficiently rescued by infecting a susceptible cell line MDBK after transient transfection of HEK293F suspension cells. According to the method provided by the invention, the BHV-1 virus seed bank can be stably obtained, the subsequent large-scale production process of BHV-1 genetic engineering vaccines is facilitated, and the production cost can be effectively reduced.

The invention relates to a recombinant cosmile of a duck adenovirus B2 reverse genetic operating system and application of the recombinant cosmile, the recombinant cosmile is a recombinant cosmile BG18FOS-2 and is constructed by using duck adenovirus B2 DAdV-B2 / delta ORF55-ORF19, the duck adenovirus B2 DAdV-B2 / delta ORF55-ORF19 is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: V202568. Compared with a duck adenovirus B2 strain BGMH, the recombinant cosmid BG18FOS-2 sequence has the differences as shown in SEQ ID No.1-2, a deletion region located between a gene ORF20 and a gene ORF53 and the like. The recombinant cosmile can successfully rescue a virus with the same biological characteristics as the duck adenovirus B2 DAdV-B2 / delta ORF55-ORF19, and a reverse genetic operating system containing the recombinant cosmile is successfully constructed.

The invention belongs to the technical field of veterinary medicine, and relates to a recombinant pseudorabies virus strain for expressing porcine acute diarrhea syndrome coronavirus S protein as well as a construction method and application of the recombinant pseudorabies virus strain. The recombinant pseudorabies virus strain is obtained by rescuing recombinant pseudorabies virus infectious clone for expressing S protein of porcine acute diarrhea syndrome coronavirus; the recombinant pseudorabies virus infectious clone comprises five Fosmid cosmiles, the five Fosmid cosmiles contain gene segments, namely Fos-1, Fos-2-dTK, Fos-3-dTK, Fos-4 and Fos-5-SADS-S, which are used for rescuing the pseudorabies virus, are mutually overlapped and cover a pseudorabies virus genome, and the Fos-5-SADS-S contains a porcine acute diarrhea syndrome coronavirus S protein gene expression cassette. After a mouse is inoculated with the recombinant pseudorabies virus strain, remarkable PRV and SADS-CoV specific antibodies and neutralizing antibodies can be induced to be generated. An immunized mouse can resist the lethal dose attack of a PRV virulent strain, and a suckling mouse produced by the immunized mouse can resist the lethal dose attack of SADS-CoV.