Nucleotide sequence, fiber2 protein, expression method of fiber2 protein and duck type 3 adenovirus and duck tembusu virus bivalent inactivated vaccine

Abstract

The invention provides a nucleotide sequence, a fiber2 protein, an expression method of the fiber2 protein and a duck type 3 adenovirus and duck tembusu virus bivalent inactivated vaccine. The nucleotide sequence is used for coding the duck type 3 adenovirus antigenic fragment fiber2 protein, the nucleotide sequence is shown as SEQ NO:2, and the amino acid sequence of the duck type 3 adenovirus antigenic fragment fiber2 protein is shown as SEQ NO:3; a fiber2 protein gene segment and a pET-32a vector segment are connected through T4DNA ligase to obtain a pET-32a-fiber2 recombinant expression vector, then the pET-32a-fiber2 recombinant expression vector is converted into an expression strain E.coli BL-21 (DE3) competent cell for cultivation, and the fiber2 protein and the duck type 3 adenovirus and duck tembusu virus bivalent inactivated vaccine are obtained from culture; used antigens of the bivalent inactivated vaccine comprise the duck type 3 adenovirus antigenic fragment fiber2 protein and inactivated duck tembusu viruses; and the volume ratio of the two antigens in the bivalent inactivated vaccine is 1:1, the duck tembusu viruses are duck tembusu virus DF2 strains, and the preservation number of the duck tembusu virus DF2 strain is CCTCC NO: V201635.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a nucleotide sequence, a fiber2 protein and an expression method, and a double inactivated vaccine of duck type 3 adenovirus and duck Tembusu virus. Background technique [0002] At present, there is only a single cell vaccine related to duck type 3 adenovirus (DAdV) vaccine, and duck Tembusu virus vaccine has only a single vaccine; as the duck type 3 adenovirus vaccine announced in patent CN108330108A, it is produced in primary duck embryo fibroblast (DEF Duck adenovirus type 3 and duck adenovirus type 2 GR strains have the highest homology, because it has been reported that the isolates of duck adenovirus type II can inactivate in primary duck embryo fibroblasts Proliferation, causing obvious cell lesions, so the virus liquid is harvested in the expanded culture of the passable duck embryo fibroblasts to obtain the virus liquid with the best virus toxicity. The preparati...

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Problems solved by technology

[0002] At present, there is only a single cell vaccine related to duck type 3 adenovirus (DAdV) vaccine, and duck Tembusu virus vaccine has only a single vaccine; as the duck type 3 adenovirus vaccine announced in patent CN108330108A, it is produced in primary duck embryo fibroblast (DEF Duck adenovirus type 3 and duck adenovirus type 2 GR strains have the highest homology, because it has been reported that the isolates of duck adenovirus type II can inactivate in primary duck embryo fibroblasts Proliferation, causing obvious cell lesions, so the virus liquid is harvested in the expanded culture of the passable duck embryo fibroblasts to obtain the virus liquid with the best virus toxicity. The preparation of primary DEF cells is complicated, easy to pollute, and costly; and At present, there are only studies and immune efficacy evaluations on duck adenovirus type 3 and duck Tembusu virus single vaccines, such as "Duck Adenovirus Type 3 Genome Sequence Analysis and Pathogenicity Research" and "Duck Tembusu Virus Inactivated Vaccine The introduction of "Development" can prepare viruses with a concentration of 10 5.52 TCID50 of duck adenovirus type 3 vaccine and 10 6.0 TCID50 (1-fold concentration) inactivated duck Tembusu virus vaccine, due to the high cost of using concentration technology to prepare vaccines with high antigen content, and the high dispersion of antibody titers in a single immunization of inactivated vaccines, the two vaccines are in the During use, ducks need to be immunized twice, and better immune protection can be achieved by increasing the number of immunizations. Repeated catching and immunization of ducks can easily cause duck stress, resulting in stress death of ducklings and egg breeding. Duck egg production drops sharply

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