Sequence analysis, expression, and binding activity of recombinant major outer sheath protein (Msp) of Treponema denticola.

摘要: The gene encoding the major outer sheath protein (Msp) of oral spirochete Treponema denticola ATCC 35405 was cloned, sequenced, and expressed in Escherichia coli. Preliminary sequence analysis showed that 5' end msp not present on 5.5-kb cloned fragment described a recent study (M. Haapasalo, K. H. Muller, V. J. Uitto, W. Leung, B. C. McBride, Infect. Immun. 60:2058-2065,1992). obtained by PCR amplification from T. genomic library, an open reading frame 1,629 bp identified as coding region for Msp combining overlapping sequences. deduced peptide consisted 543 amino acids had molecular mass 58,233 Da. typical prokaryotic signal with potential cleavage site peptidase 1. Northern (RNA) blot showing transcript to be approximately 1.7 kb consistent identification promoter consensus located optimally upstream transcription termination found downstream stop codon. entire amplified DNA E. coli using tightly regulated T7 RNA polymerase vector system. Expression toxic when present. High levels were produced inclusion bodies putative deleted replaced vector-encoded sequence. Recombinant purified homogeneity clone containing full-length adhered immobilized laminin fibronectin but bovine serum albumin. Attachment recombinant decreased presence soluble substrate. increased pretreatment substrate Msp. These studies lend further support hypothesis mediates extracellular matrix binding activity denticola.