Cloning and purification of recombinant proteins of Mycoplasma hyopneumoniae expressed in Escherichia coli

Mycoplasma hyopneumoniae, the etiological agent of swine enzootic pneumonia, is an important pathogen in the swine industry worldwide. Vaccination is the most cost-effective strategy for controlling and prevention of this disease. However, investigations on pathogenicity mechanisms as well as curren...

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Vydáno v:Protein expression and purification Ročník 69; číslo 2; s. 132 - 136
Hlavní autoři: Simionatto, Simone, Marchioro, Silvana B., Galli, Vanessa, Hartwig, Daiane D., Carlessi, Rodrigo M., Munari, Fernanda M., Laurino, Jomar P., Conceição, Fabricio R., Dellagostin, Odir A.
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States Elsevier Inc 01.02.2010
Témata:
Animals
Antigens, Bacterial - immunology
Bacterial Proteins - genetics
Bacterial Proteins - immunology
Bacterial Vaccines - genetics
Bacterial Vaccines - immunology
Cloning, Molecular
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Mycoplasma hyopneumoniae
Mycoplasma hyopneumoniae - genetics
Mycoplasma hyopneumoniae - metabolism
Pneumonia of Swine, Mycoplasmal - microbiology
Pneumonia of Swine, Mycoplasmal - prevention & control
Protein purification
Recombinant protein
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Swine
Vaccines, Subunit - genetics
Vaccines, Subunit - immunology
Vaccines, Synthetic - genetics
Vaccines, Synthetic - immunology
Mycoplasma hyopneumoniae
Protein purification
Recombinant protein
ISSN:1046-5928, 1096-0279, 1096-0279
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Shrnutí:Mycoplasma hyopneumoniae, the etiological agent of swine enzootic pneumonia, is an important pathogen in the swine industry worldwide. Vaccination is the most cost-effective strategy for controlling and prevention of this disease. However, investigations on pathogenicity mechanisms as well as current serological detection methods and the development of new recombinant subunit vaccines are hampered by the lack of known and well characterized species-specific M. hyopneumoniae antigens. In this work, 54 predicted genes encoding proteins with potential to be used as subunit vaccine or antigens in diagnostic tests were selected, amplified by PCR and cloned into Escherichia coli expression vectors. Recombinant protein expression, solubility and yields were analyzed. The majority of the recombinant proteins were expressed in inclusion bodies. After solubilization with urea or N-lauroyl sarcosine, recombinant proteins were purified by Ni 2+ affinity chromatography. This approach allowed purification of thirty recombinant M. hyopneumoniae proteins which will be evaluated as vaccine candidates and/or as antigens to be used in diagnostic tests.
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ISSN:1046-5928
1096-0279
1096-0279
DOI:10.1016/j.pep.2009.09.001