9 results about "Cosmid" patented technology

The invention discloses a recombinant herpesvirus turkey strain for co-expressing an H9 subtype AIVHA gene and ILTVgB, gD and gI genes as well as a construction method and application of the recombinant herpesvirus turkey strain, and belongs to the technical field of medicine or veterinary medicine. The recombinant herpesvirus of turkeys is obtained by co-transfecting CEF cells with recombinant clay containing H9 subtype AIVHA genes and ILTVgB genes, recombinant clay containing ILTVgD and gI genes and parent clay containing HVT genome DNA fragments and then carrying out virus rescue. The recombinant herpesvirus of turkeys is prepared by the following steps: cotransfecting CEF cells with recombinant clay containing H9 subtype AIVHA genes and ILTVgB genes, recombinant clay containing ILTVgD and gI genes, and parent clay containing HVT genome DNA fragments. The recombinant virus strain obtained by the invention has good in-vitro replication ability and genetic stability. After SPF chickens are immunized, the SPF chickens can obtain immune protection against H9 subtype AIV and ILTV virulent viruses. The invention provides a new technical means for simultaneously preventing the H9 subtype avian influenza and the avian infectious laryngotracheitis.

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 discloses a recombinant herpesvirus turkey strain for co-expressing an H9 subtype AIVHA gene, an IBDVVP2 gene and ILTVgB, gD and gI genes as well as a construction method and application of the recombinant herpesvirus turkey strain, and belongs to the technical field of medicine or veterinary medicine. The recombinant herpesvirus of turkeys is obtained by co-transfecting CEF cells with recombinant clay containing H9 subtype AIVHA genes and ILTVgB genes, recombinant clay containing IBDVVP2 genes, recombinant clay containing ILTVgD and gI genes and parent clay containing HVT genome DNA fragments, and then carrying out virus rescue. The recombinant herpesvirus of turkeys is obtained by the following steps: cotransfecting CEF cells with recombinant clay containing H9 subtype AIVHA genes and ILTVgB genes, recombinant clay containing IBDVVP2 genes, recombinant clay containing ILTVgD and gI genes, and parent clay containing HVT genome DNA fragments. The recombinant virus strain obtained by the invention has good in-vitro replication ability and genetic stability. After the SPF chicken is immunized, the SPF chicken can obtain strong immune protection on H9 subtype AIV (avian influenza virus), IBDV (infectious bursal disease virus) and ILTV (Illuminal Low Temperature Virus). The invention provides a new technical means for simultaneously preventing the H9 subtype avian influenza, the infectious bursal disease and the infectious laryngotracheitis.

The invention provides a recombinant Marek's disease virus strain for co-expressing a Newcastle disease virus F gene and an H9 subtype avian influenza virus HA gene and application, and belongs to the field of genetic engineering vaccines. According to the invention, by using a recombinant cloning technology, an expression framework containing a Newcastle disease virus gene VII type strain F gene and an H9 subtype avian influenza virus HA gene is inserted into a spacer region between Marek's disease virus genome UL45 and UL46 genes; according to the construction method, the recombinant cosmid is obtained by inserting a Newcastle disease virus F gene and an H9 subtype avian influenza virus HA gene expression framework between Marek's disease virus genome UL45 and UL46 genes, and the recombinant virus for co-expressing the Newcastle disease virus F gene and the H9 subtype avian influenza virus HA gene is obtained by rescue of the recombinant cosmid. The vaccine prepared from the recombinant Marek's disease virus strain provided by the invention can prevent Newcastle disease, H9 subtype avian influenza and Marek's disease at the same time, and has a good market application prospect.

The invention discloses a recombinant Marek's virus strain for co-expressing an H9 subtype AIV HA gene and an IBV S gene as well as a construction method and application of the recombinant Marek's virus strain, and belongs to the technical field of medicine or veterinary medicine. The recombinant Marek's virus is obtained by co-transfecting CEF cells with recombinant coplasmids containing HA gene and S gene expression cassettes and recombinant coplasmids containing MDV genome DNA fragments and then carrying out virus rescue. Specifically, by using a recombinant cloning technology, an expression cassette containing an H9 subtype AIV HA gene and an IBV S gene is inserted between 105561-105563 nucleotides of a serum type 1 MDV attenuated live vaccine CVI988 strain, so as to construct and obtain a recombinant clone in which the HA gene and the S gene are inserted in an MDV genome, and the recombinant clone rescues to obtain a recombinant MDV vaccine strain C20301 for co-expressing the AIV HA gene and the IBV S gene. The invention also relates to application of the recombinant MDV vaccine strain in preparation of vaccines for simultaneously preventing H9 subtype avian influenza, infectious bronchitis and Marek's disease.

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.