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Attenuated Actinobacillus pleuropneumoniae as a bacterial vector for expression of Mycoplasma hyopneumoniae P36 gene
Author(s) -
Zou HaoYong,
Liu XinJun,
Ma FengYing,
Chen Pin,
Zhou Rui,
He QiGai
Publication year - 2011
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1556
Subject(s) - mycoplasma hyopneumoniae , actinobacillus pleuropneumoniae , microbiology and biotechnology , virulence , biology , attenuated vaccine , mutant , antibody , antigen , pasteurellaceae , virology , serotype , gene , haemophilus influenzae , immunology , genetics , antibiotics
Background Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae are causative agents of porcine pneumonia. Over the last few years, attenuated A. pleuropneumoniae live vaccines have been shown to provide protection against A. pleuropneumoniae infection. We postulated that attenuated A. pleuropneumoniae could additionally be used as a vaccine vector for protection against M. hyopneumoniae . Methods A mutant strain of A. pleuropneumoniae , SLW36, was constructed by replacing the urease structural gene of mutant strain SLW03 of A. pleuropneumoniae with the L‐lactate dehydrogenase gene (p36) of M. hyopneumoniae by transconjugation and counter selection. The urease function and the growth kinetics of SLW36 were measured. Protein expression of P36 was analyzed by sodium dodecylsulfate‐polyacrylamide gel electrophoresis and western blotting. The attenuated virulence and immunity of SLW36 were analyzed in a mouse model. Results The mutant strain SLW36 was urease negative and four‐fold less virulent than the parental strain SLW03. There were no differences in expression levels of p36 at different culture time‐points and the foreign gene was stable after in vitro passage. Immunoglobulin G responses against p36 antigen and M. hyopneumoniae whole‐cell antigen were detected. Conclusions The mutant strain SLW36 can induce antibody against p36 and M. hyopneumoniae. The mutant strain SLW36 has the potential to be used as a live vaccine for protection against A. pleuropneumoniae and M. hyopneumoniae . Studies in pigs are needed to confirm protective levels of antibodies and to check for rare side‐effects of the vaccine. Copyright © 2011 John Wiley & Sons, Ltd.