Construction and expression of lactic acid bacteria for recombination of GP5 gene of porcine reproductive and respiratory syndrome virus

Abstract

The invention discloses construction and expression of lactic acid bacteria for recombination of a GP5 gene of a porcine reproductive and respiratory syndrome virus and belongs to the technical field of biology. The construction and expression are characterized by comprising the following steps of cloning and sequencing the GP5 gene of the porcine reproductive and respiratory syndrome virus, carrying out dual-enzyme digestion on a recombinant plasmid pMD19(simple)-dGP5 and a carrier pNZ8149 by Nco I and Xba I, connecting purified enzyme-digested products by T4DNA ligases; carrying out electrotransformation on the connected products into competence NZ3900 of lacF gene deletion type lactic acid bacteria, culturing on an LM17 recovery medium, and obtaining recombinant expression plasmid pNZ8149-dGP5 by Ellike plate screening. The construction and expression disclosed by the invention have the beneficial effects that the GP5 gene of the porcine reproductive and respiratory syndrome virus can be recombined in the expression carrier pNZ8149 of a lactococcus lactis; proved by detection, the exogenous gene GP5 can be expressed in the lactic acid bacteria by the carrier pNZ8149 and has reactogenicity, so that an experimental basis is provided for next-step development of safe-effective-cheap oral preparations.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to the field of biotechnology of the construction and expression of a porcine reproductive and respiratory syndrome virus GP5 gene recombinant lactic acid bacteria. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which can cause reproductive dysfunction in sows and Severe respiratory diseases in pigs have brought huge economic losses to my country's pig industry. Porcine reproductive and respiratory syndrome virus particles are spherical, with a diameter of 48-83 nm and an envelope. PRRSV is a single-stranded, positive-strand RNA virus with a genome length of about 15kb, a cap structure at the 5' end, and a poly(A) tail of polyadenylation after the non-coding region at the 3' end. It encodes 9 ORFs, the genes are ORF1a, ORF1b, ORF2a, ORF2b,...

AI Technical Summary

Problems solved by technology

It is generally believed that the immune effect of the attenuated vaccine is better than that of the inactivated vaccine, which can reproduce in cells and stimulate the body to produce humoral immunity and cellular immunity at the same time, but there may be risks of incomplete inactivation and strong virulence

Inactivated vaccines cannot replicate in cells, the immune response is not strong, and multiple immunizations are required to achieve effective antibody levels, and the immune effect is not ideal

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