Genetic engineering vaccine of lymphocystis disease of flounder paralichthys olivaceus and production method thereof

Abstract

The invention discloses a genetic engineering vaccine of lymphocystis disease of flounder paralichthys olivaceus and a production method thereof, which is characterized in that the method for producing the genetic engineering vaccine comprises the following steps: inserting gene MCP segments (size: 0.6 KB) of nucleocapsid proteins of lymphocystis disease virus (LCDV) into an eukaryotic expression vector pEGFP-N2 so as to form recombinant plasmids pEGFP-N2-LCDV-MCP (size: 0.6 KB); translating the recombinant plasmids into escherichia coli so as to prepare production strains, lining the production strains to a Mi nca agar plate so as to separate and purify the production strains and then prepare and determine qualified level-1 strains; then inoculating the level-1 strains in an agar flat bottle, after cultivating the level-1 strains for 24h-48h at a temperature of 37 DEG C, identifying the obtained level-1 strains and determining qualified level-2 strains; carrying out fermentation and cultivation on the obtained level-2 strains so as to obtain fermentative level-2 strain culture solution; carrying out bacteria collection and centrifugation as well as plasmid DNA extraction and purification on the fermentative level-2 strain culture bacterium solution, then carrying out collection, centrifugation, plasmid DNA extraction, purification and routine inspection on the cultured bacterium solution, if qualified, the genetic engineering vaccine is obtained. The genetic engineering vaccine of the invention has the advantages of complete and simple production process, good stability and low cost; and compared with current inactivated vaccines, attenuated vaccines, polypeptide vaccines, subunit vaccines and recombinant polypeptide vaccines and the like, the genetic engineering vaccine of the invention has good immunization effect and no adverse effect.

Description

technical field [0001] The invention relates to a seawater fish virus disease genetic engineering vaccine and a production method thereof, in particular to a flounder flounder lymphocyst virus disease genetic engineering vaccine and a production method thereof. Background technique [0002] Lymphocystic disease is a viral infectious disease caused by lymphocyst virus. It has the characteristics of both horizontal and vertical transmission and can infect a variety of fish. It is widely spread in the important and valuable fish species such as flounder, red sea bream, grouper, perch, etc., which are important aquaculture economic fish in my country, causing serious economic losses. At present, the control of mariculture diseases in my country is still dominated by chemical drugs such as antibiotics. The abuse of drugs has led to drug residues in aquatic products and seriously affected food safety. In addition, the extensive use of antibiotics and chemicals has also caused the ...

AI Technical Summary

Problems solved by technology

Genetic engineering vaccines are to clone foreign genes into eukaryotic plasmid expression vectors, and then inject recombinant plasmid DNA into animals. When foreign genes enter and express in living cells, they can induce both humoral immune responses and long-term immune responses. The immune memory of the vaccine is known as the third generation vaccine, and the production and processing technology of large-scale preparation of clinical-grade plasmid DNA has gradually attracted attention. They are not mature, and there is no relevant quality assurance system, which greatly hinders the development of the industry