ILTV gD glycoprotein nucleotide sequence and amino acid sequence, recombined virus bacterin thereof and application of the bacterin

A nucleotide sequence and glycoprotein technology, applied in the field of virus vaccines, can solve the problems of long culture time, strong virulence, and time-consuming and labor-intensive vaccination methods.

Inactive Publication Date: 2008-07-02
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the invention patent is to overcome the attenuated vaccine that can cause latent infection, and latently infected chickens will be infected for life and become a new source of infection; secondly, the attenuated vaccine virus can be transmitted from vaccinated chickens to non-vaccinated chickens, and spread between chickens During the process, the virulence may become stronger; and in the production process of the attenuated chicken infectious laryngotracheitis vaccine, the virus multiplication is mainly inoculated with the allantoic membrane of chicken embryos. This inoculation method is time-consuming and laborious, and the cultivation time is long After inoculation, it needs to continue to cultivate for about 6 days to collect the virus, and the amount of virus harvested is small, only the thickened allantoic membrane; and other shortcomings, providing the same immune effect as the existing ILTV attenuated vaccine, and at the same time can overcome the attenuated virus The new vaccine with the disadvantage of the vaccine will lay the foundation for the prevention and control of ILT and the ultimate eradication of ILT

Method used

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  • ILTV gD glycoprotein nucleotide sequence and amino acid sequence, recombined virus bacterin thereof and application of the bacterin
  • ILTV gD glycoprotein nucleotide sequence and amino acid sequence, recombined virus bacterin thereof and application of the bacterin
  • ILTV gD glycoprotein nucleotide sequence and amino acid sequence, recombined virus bacterin thereof and application of the bacterin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Isolation and identification of ILTV Henan Changge strain and its gC and gD gene cloning and sequencing

[0091] Abstract: Through the virus isolation and identification of diseased chicken throat and trachea from a chicken farm in Changge, Henan, an ILTV Henan Changge strain was obtained; the full gC and gD genes of ILTV Henan Changge strain were amplified and cloned Sequencing; after analysis, its nucleotide sequence and amino acid sequence have high homology with the ILTV gC, gD nucleotide sequence and amino acid sequence read by GenBank, both of which are between 99.3% and 99.9%, indicating that ILTV gC, gD The genes are relatively conservative, and ILTV has very few antigenic gene mutations. Therefore, an effective vaccine can protect all ILTV virulent strains from attack and make local eradication of ILT a reality.

[0092] Keywords: chicken infectious laryngotracheitis virus; gC gene; gD gene; cloning

[0093] Infectious laryngotracheitis (ILT) is an acute upper resp...

Embodiment 2

[0136] The expression of ILTV Henan Changge strain gD gene in recombinant poxvirus and its immunological efficacy test report Abstract: The gD gene of infectious laryngotracheitis virus (ILTV) glycoprotein cloned into pGEM-T easy, through EcoR I enzyme The cutting site was subcloned to the downstream of the LP2EP2 promoter of the pSY538 vector to construct the pSY538-gD vector; Pst I and XbaI were used to cut the E. coli LacZ gene under the control of the poxvirus promoter in the pSC11 vector, and after filling in with DNA polymerase, The pSY538-gD vector is connected to the Sma I restriction site to construct the pSY538-gD-LacZ vector; the large fragments of gD and LacZ are digested with Not I and then connected to the vector pSY681 Not I restriction site to construct pSY681-gD- LacZ carrier. The pSY538-gD-LacZ plasmid was transfected into poxvirus (FPV) infected CEF cells by liposome transfection method; the recombinant virus was screened and purified by blue spot screening test...

Embodiment 3

[0181] Animal test report

[0182] Study on Immune Efficacy of ILTV Henan Changge Strain gD Gene Recombinant Fowlpox Virus Vaccine

[0183] Abstract: In the test, ILTV Henan Changge strain gD gene recombinant fowlpox virus vaccine was inoculated subcutaneously in the inner avascular part of the wing, and 35-day-old chickens were immunized. The serum neutralizing antibody test showed that the ILTV gD neutralization test in the serum of chickens after the recombinant virus immunization test The half-protected dose (PD50) of the serum was 1:55; and the PD50 of the chickenpox antibody in the serum was 1:49. The experimental chicken challenge test showed that the protection rate of the immunized group was 96% (24 / 25), the morbidity rate of the control group was 92% (23 / 25), and the mortality rate was 40% (10). / 25). The test results show that the ILTV Henan Changge strain gD gene recombinant fowlpox virus vaccine is a vaccine strain with small side effects and good immunity.

[0184] K...

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Abstract

The invention provides a novel vaccine with the same immunoefficiency as the prior infectious laryngotracheitis virus (ILTV) attenuated vaccine and capable of overcoming dormant infection induced by the attenuated vaccine. The invention provides the nucleotide sequence and amino acid sequence of an ILTV glycoprotein D (gD) and a preparation method, as well as a recombinant vaccine containing a viral vector (fowlpox virus vector) and ILTV gD. The fowlpox virus is obtained by homologous arm recombination of ILTV gD. The invention also provides the preparation method of the recombinant vaccine. The invention clones gD gene of ILTV Henan isolate, and performs animal experiments of the immunoprotection of the recombinant fowlpox virus. The novel vaccine has the same immunoefficiency as the prior ILTV attenuated vaccine and is capable of overcoming the shortcomings of the attenuated vaccine. The invention is convenient for large-scale production, and provides a foundation for prevention and elimination of infectious laryngotracheitis (ILT).

Description

Technical field [0001] The invention belongs to the field of recombinant animal vaccines, and specifically relates to an ILTV gD glycoprotein nucleotide sequence and amino acid sequence and a recombinant virus vaccine and application of the vaccine. technical background [0002] The prevalence of infectious laryngotracheitis in chickens and the situation of vaccine immunity [0003] Infectious laryngotracheitis (ILT) is an acute respiratory infectious disease of chickens caused by the avian infectious laryngotracheitis virus (ILTV). The disease spreads rapidly, has a high incidence, the infection rate is 80-90%, and the mortality rate is 20. -30%, sometimes as high as 70%, can reduce the egg production rate of chickens by 15-60% after the onset of disease. The disease is endemic in many countries and places. According to reports, it was first discovered in the United States in 1925. At present, more than 40 countries in Europe, the United States, Asia, Australia and other contine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/33C07K14/005C12N15/09C12N15/10A61K48/00A61K39/12C12N15/863
Inventor 杨明凡崔保安陈红英苗舜尧
Owner HENAN AGRICULTURAL UNIVERSITY
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