Recombinant adenovirus and construction method thereof

Abstract

The invention provides a recombinant adenovirus and a construction method thereof. The construction method comprises the following steps of: inserting swine fever virus E2 genes into a multiple cloning site Bg1 II and a Sa1 I enzyme cutting site on the downstream of a CMV (cytomegalovirus) promotor of the adenovirus; and inserting porcine reproductive and respiratory syndrome virus GP5 genes into enzyme cutting sites Kpn I and Not I on the downstream of the CMV promotor of the adenovirus. The problems of low expression yield and weak protection effect when the E2 and GP5 genes are fused and connected in series are solved. Only by once immune vaccination, a pig can obtain antibodies for a swine fever virus and a porcine reproductive and respiratory syndrome virus. The recombinant adenovirus is a nonreplicative virus and is not reproduced in a pig body. The phenomena of virus atavism and toxicity strengthening caused by long-term vaccination of an attenuated vaccine are avoided. Moreover, the recombinant adenovirus only contains main protection genes of the swine fever virus and the porcine reproductive and respiratory syndrome virus and does not contain other genes of viruses, so that recombination cannot occur between the attenuated vaccine and a wild strain. A vaccine prepared from the recombinant adenovirus has a protection effect of 87.5 percent on coalition attack of the swine fever virus and the porcine reproductive and respiratory syndrome virus after used for immunizing an animal. The problem that the swine fever and porcine reproductive and respiratory syndrome vaccines cannot be immunized simultaneously is solved. The problems of strengthened toxicity and homologous recombination after the immunization of the attenuated vaccine are also solved.

Description

technical field [0001] The invention relates to a recombinant adenovirus, in particular to a recombinant adenovirus capable of separately expressing the E2 gene of swine fever virus and the GP5 gene of PRRS virus in the same adenovirus and a construction method thereof, belonging to the field of biotechnology. Background technique [0002] Swine fever is one of the main infectious diseases that harm the pig industry. At present, immunization is still the main measure for the prevention and treatment of swine fever, and it is a necessary means to eradicate the disease and prevent the spread of wild virus to pigs without swine fever. No matter in the past, at present or in the future, swine fever vaccine plays an extremely important role in controlling and eliminating swine fever. Scholars from various countries have never given up on the research and development of swine fever vaccine. Traditional swine fever vaccines include inactivated vaccines and attenuated vaccines, but...

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Problems solved by technology

[0007] In order to solve the problems that the attenuated swine fever vaccine and the attenuated PRRS vaccine cannot be immunized at the same time, and the phenomenon of virus reversion and independent enhancement caused by long-term inoculation of the attenuated vaccine, and the recombination of the attenuated vaccine strain and the wild virus, the invention provides a Recombinant adenovirus expressing classical swine fever virus E2 gene and PRRS virus GP5 gene in the same adenovirus and its construction method

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