Preparation method of envelope protein A gene vaccine for fowl cholera

Abstract

The invention discloses a preparation method of a nucleic acid vaccine for fowl cholera, which comprises the following steps of: designing primers by a fowl cholera bacterium genome sequence; carrying out PCR (polymerase chain reaction) amplification by taking a fowl cholera bacterium CVCC474 strain genome as a template to obtain a target gene; carrying out enzyme digestion on the target gene andan eukaryotic expression vector pcDNA3.1(+) by utilizing a restriction enzyme KpnI and a restriction enzyme EcoRI; then connecting the target gene with the eukaryotic expression vector pcDNA3.1(+); converting into the Escherichia coli competence JM83; extracting plasmids; and carrying out enzyme digestion identification to obtain the nucleic acid vaccine for fowl cholera. The animal experiment detection shows that the nucleic acid vaccine for fowl cholera prepared by the method disclosed by the invention can reduce the occurrence of fowl cholera and can reduce the intrusion of fowl cholera bacteria on fowls; and the preparation method of the nucleic acid vaccine is simple and easy to operate.

Description

technical field [0001] The invention relates to a preparation method of a bacterial disease vaccine, in particular to a preparation method of fowl cholera outer membrane protein A gene vaccine. Background technique [0002] Fowl cholera, also known as poultry Pasteurella disease, is a contagious disease of poultry caused by fowl cholera bacteria (also known as poultry Pasteurella). It is a common multiple infection in my country. The disease is distributed in all countries in the world, and almost all poultry can be infected with the disease. Its epidemic has seriously affected the healthy development of the poultry industry and brought huge economic losses to the poultry industry. [0003] At present, the control of fowl cholera in my country is mainly treated with antibiotic drugs, but there are certain shortcomings in drug therapy, and it is easy to relapse after drug withdrawal. Long-term application may have obvious toxic effects on poultry, causing the egg production ra...

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Problems solved by technology

[0003] At present, the control of fowl cholera in my country is mainly treated with antibiotic drugs, but there are certain shortcomings in drug therapy, and it is easy to relapse after drug withdrawal. Long-term application may have obvious toxic effects on poultry, causing the egg production rate of laying hens to decline. Moreover, long-term or repeated medication can easily lead to drug resistance of fowl cholera bacteria

[0004] At present, the main preventive measures for fowl cholera are vaccine injections. Commonly used vaccines include attenuated live vaccines and inactivated vaccines. However, the immune effects of the two vaccines are not particularly ideal. The immune protection is not high enough, and there are certain side effects after injecting poultry, which may even cause some deaths

Inactivated vaccines have the problems of low protection rate and short immunity period, which are more serious than attenuated live vaccines, so the immune effect of inactivated vaccines is not as good as that of attenuated live vaccines