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Preparation method and applications of chicken Newcastle disease and egg drop syndrome bivalent genetic engineering subunit vaccine

A technology of egg drop syndrome and subunit vaccines, applied in the field of genetic engineering veterinary vaccines, can solve unseen problems and achieve the effects of promoting development, good social and economic benefits, and promoting social and economic progress

Inactive Publication Date: 2018-09-25
成都金洛克锶生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no combined subunit vaccine composed of protective antigens of Newcastle disease and egg drop syndrome virus

Method used

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  • Preparation method and applications of chicken Newcastle disease and egg drop syndrome bivalent genetic engineering subunit vaccine
  • Preparation method and applications of chicken Newcastle disease and egg drop syndrome bivalent genetic engineering subunit vaccine
  • Preparation method and applications of chicken Newcastle disease and egg drop syndrome bivalent genetic engineering subunit vaccine

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Preparation of embodiment 1 fusion protein

[0021] 1. Construction of HN-knob protein particles

[0022] 1.1 Gene and primer synthesis

[0023] The expression sequence was recombined into the expression vector by the synthetic gene described below. The gene and primers were synthesized by Jinweizhi Company, and the synthetic gene sequence was recombined into the plasmid vector pUC19, and the plasmids containing the target gene were obtained as pUC-Hk; the following is the sequence of the synthetic gene fragment.

[0024] Synthetic gene fragment Hk:

[0025]CCTAGGGCCACCATGGCCTGGATGATGCTTCTCCTCGGACTCCTTGCTTATGGATCAGGAGTCGACTCTACCCCTCTCACCAGGATCATCTCCATGGGCAACAACCTCTTCGACAGCGGCTACGAAATTTTTGCCAGCTGCCCCCAGAACAAGGCCGCCAAGGTGGCTGGCTACGTGTACCTGACATCCGTGGGAGGCCTGGTCCACGGCACCATCCAAATCAAGGCCACCGCCGGCTACTGGTTTACCGGCGGCAACAGCGTGCAGGAGTCCATCAGGTTCGGCCTGGTGCTGTGTCCCTTCAGCGCCCGGGACCCTACAGCTAATCTGAGCGGCTGGCCCGCTCCTGTGGTGTGGAGCGGAGACTCCAACACCCCCCTGTACTTCGCCGCCAACGCCATCAGCTACACCAACAACC...

Embodiment 2

[0042] Example 2 Detection of immune efficacy

[0043] 1. Immune potency test of recombinant dual antigen protein

[0044] The dual antigen protein HN-knob obtained in Example 1 was mixed with an adjuvant (chitosan) at a ratio of 1:1 to make a subunit vaccine, and the immune efficacy test was carried out. Eighty 21-day-old SPF chickens were randomly divided into 4 groups: the blank control group and the dual antigen protein HN-knob vaccine group (0.5 mg / feather). Blood was collected from the hindwing vein at the 2nd, 4th, and 6th week after inoculation, and the serum was separated to measure the antibody titer by HI (hemocoagulase inhibition test method). The specific method is: take the test serum and serially dilute it with PBS, then take 0.025ml each and add it to 4 units of HA (Hemagglutinin, hemagglutinin) antigen in an equal volume. After 20 minutes, 0.05ml of 0.5% chicken erythrocyte suspension was added and mixed respectively. After incubation at room temperature for...

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Abstract

The invention provides a chicken Newcastle disease and egg drop syndrome bivalent genetic engineering subunit vaccine and uses thereof, and a preparation method thereof, wherein the preparation methodcomprises: fusing the HN fragment of chicken Newcastle disease virus and the knob fragment of egg drop syndrome virus through a linking peptide to form a vector, and expressing in mammalian cells. According to the present invention, with the matched application of the obtained genetic engineering subunit vaccine and the adjuvant to immunize chicken, the chicken Newcastle disease and egg drop syndrome bivalent genetic engineering subunit vaccine has good immunogenicity, can be safely applied to poultry, and can achieve good prevention effects.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering veterinary vaccines, in particular to a dual genetic engineering subunit vaccine of Newcastle disease and egg drop syndrome and a preparation method thereof. Background technique [0002] Newcastle disease (ND) is a contagious disease mainly infecting poultry and wild poultry caused by Newcastle disease virus (NDS). Once an outbreak occurs, it will bring a devastating blow to the poultry farming industry. The International Veterinary Bureau (OIE ) has listed it as a category A disease that must be reported. The incidence of ND in my country is very high, and the mortality rate is as high as more than 90%, which seriously endangers the healthy development of my country's chicken breeding industry. [0003] Egg drop syndrome (EDS) is an infectious disease caused by egg drop syndrome virus (EDSV). The main performance of the chicken flock, and lasts for a long time, brings huge economic...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/17A61K39/235C12N15/85C12N15/62A61P31/14A61P31/20
CPCA61K39/12A61K2039/552A61K2039/70C07K14/005C07K2319/00C12N15/62C12N15/85C12N2710/10222C12N2710/10234C12N2760/18122C12N2760/18134
Inventor 李生伟付伟张晟代燕平王雯茜何刚程琳高小平
Owner 成都金洛克锶生物技术有限公司
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