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FEMS Microbiol Lett ; 183(1): 15-21, 2000 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-10650196

RESUMO

The gap gene encodes the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The gene was cloned and sequenced from the Mycoplasma hominis type strain PG21(T). The intraspecies variability was investigated by inspection of restriction fragment length polymorphism (RFLP) patterns after polymerase chain reaction (PCR) amplification of the gap gene from 15 strains and furthermore by sequencing of part of the gene in eight strains. The M. hominis gap gene was found to vary more than the Escherichia coli counterpart, but the variation at nucleotide level gave rise to only a few amino acid substitutions. To verify that the gene was expressed in M. hominis, a polyclonal antibody was produced and tested against whole cell protein from 15 strains. The enzyme was expressed in all strains investigated as a 36-kDa protein. All strains except type strain PG21(T) showed reaction to a 104-kDa band in addition to the expected 36-kDa band. The protein reacting at 104 kDa is a M. hominis protein with either an epitope similar to one on GAPDH, or it is an immunoglobulin binding protein.


Assuntos
Variação Genética , Gliceraldeído-3-Fosfato Desidrogenases/genética , Mycoplasma hominis/enzimologia , Mycoplasma hominis/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , Mapeamento Cromossômico , Clonagem Molecular , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Gliceraldeído-3-Fosfato Desidrogenases/química , Gliceraldeído-3-Fosfato Desidrogenases/imunologia , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Humanos , Dados de Sequência Molecular , Infecções por Mycoplasma/microbiologia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Coelhos , Análise de Sequência de DNA
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