Swine mycoplasma hyopneumoniae p52 gene recombinant adenovirus and construction method and application thereof

Abstract

The invention discloses a swine mycoplasma hyopneumoniae p52 gene recombinant adenovirus and a construction method and an application thereof. In the invention, a p52 gene is cloned into a shuttle vector pShuttle-CMV and is linearized, is electro transformed into BJ5183 fungus to carry out homologous recombination with a skeleton vector pAdEasy-1 contained therein, is mixed with liposome to transfect AD-293 cells to be packaged into complete adenoviruses. After being identified by RT-PCR and indirect immunofluorescence, virus is augmented, and fluid with the virus is collected, Balb / c mice areimmunized, and the immunity result is analyzed. Results show that recombinant adenovirus expressing the p52 gene is constructed successfully; the microorganism preservation number thereof is CGMCC NO. 2986. The recombinant adenovirus can correctly express a target protein and has high potency. Animal experiments prove that the recombinant adenovirus of the invention can generate special humoral immunity and mucosal immunization response reaction.

Description

technical field [0001] The present invention relates to a recombinant virus, in particular to a Mycoplasma hyopneumoniae p52 gene recombinant adenovirus and its construction method. The present invention also relates to the application of the recombinant adenovirus in the prevention or treatment of Mycoplasma hyopneumoniae, belonging to the prevention and control of Mycoplasma hyopneumoniae field. Background technique [0002] Mycoplasma hyopneumoniae (Mhp) is the main cause of porcine bronchopneumonia (Mycoplasmal pneumoniae of swine, Mps), also known as swine enzootic pneumonia (SwineEnzootic Pneumoniae, SEP) (ROSS R F. Disease of Swine [M] . Iowa: Iowa State University Press, 1986: 469-483.). Mhp is a contagious infectious disease unique to pigs. Once it is introduced into pigs, it can occur continuously and is difficult to eliminate. It also exists widely in all countries in the world and seriously endangers the development of pig farming. [0003] Mhp has a simple str...

AI Technical Summary

Problems solved by technology

Among the currently used vaccines, the domestic attenuated vaccine can produce effective cellular immunity, and the protection rate can reach about 80%.

Although inactivated vaccines do not require high injection techniques and less stress response, the protection rate is greatly reduced because it is difficult to stimulate effective cellular immunity and mucosal immunity (TAJIMA M, YAGIHASHIT. Isolation of Mycoplasma hyopneumoniae with porcine respiratory epithelium as observed by electron microspy [J]. Infect Immun, 1982, 37 (4): 1162-1169; Xin Jiuqing, Wang Liang, Li Yuan. Research progress of mycoplasma swine pneumonia [J]. Pig Industry Science, 2006, 34 (11) , 19~22. XIN Jiu-qing, WANG Liang, LI Yuan. The research progression of Mycoplasmal pneumoniae of swine[J]. Swine IndustryScience, 2006, 34(11), 19~22.(in Chinese))

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