Porcine reproductive and respiratory syndrome bivalence recombinant adenovirus vaccine and preparation method thereof

Abstract

The invention discloses a porcine reproductive and respiratory syndrome divalent recombination adenovirus vaccine and the preparation method thereof. The invention belongs to the technical field of biological vaccine preparation. The vaccine can be prepared by the following steps: a GP5-2A-M fusion protein gene can be constructed by inserting an FMDV2A gene with self craking between PRRSV GP5 and M protein; homologous recombination is carried out on the GP5-2A-M fusion protein gene and adenovirus backbone plasmid pAdEasy-1; recombination adenovirus rAd-GP5-2A-M is prepared by restriction enzyme and HEK-293A cells transfection, and the divalent recombination adenovirus vaccine is prepared by the technology and the steps such as purification, amplification, and the like. After expression, the aggregate protein GP5-2A-M constructed by the invention is self cracked into GP5 and M protein, as well as exerts the viral neutralization of GP5 and the immune function of the M protein; the vaccine has stable titer with the virulent valence being 10<10.43>TCID<50>/1.0ml, as well as has both the duplication characteristic of a routine attenuated vaccine and the safety of an inactivated vaccine; the divalent recombination adenovirus vaccine can be popularized in and applied to the control work of porcine reproductive and respiratory syndrome.

Description

technical field [0001] The invention belongs to the technical field of biological vaccines, in particular to a bivalent recombinant adenovirus vaccine for immune protection against porcine reproductive and respiratory syndrome and a preparation method thereof. technical background [0002] Porcine reproductive and respiratory syndrome (porcine reproductive and respiratory syndrome, PRRS) is caused by porcine reproductive and respiratory syndrome virus (PRRSV). Infectious disease characterized by high mortality. [0003] Since it was first reported in North America in 1987, it has spread to almost all countries and regions with developed swine industry in the world, causing huge economic losses to the pig industry and its product trade, and has become one of the main viral diseases in the pig industry. one. The disease broke out in my country at the end of 1995, and then spread rapidly to most provinces in the country. In recent years, the PRRS epidemic has aggravated agai...

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Problems solved by technology

However, these strategies have some defects, such as the low initial efficiency of the expressed gene, the serious imbalance between the expression of the upstream gene and the downstream gene, mutual interference between the promoters, and the nature of the expressed protein is not the same as that of the natural protein, etc.

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