Expression of the Chlamydia trachomatis major outer membrane protein-encoding gene in Escherichia coli: role of the 3′ end in mRNA stability

Ravi Kaul, Matthew J.J. Duncan, James Guest, Wanda M. Wenman

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

The major outer membrane protein (MOMP)-encoding gene (ompl) of Chlamydia trachomatis has been cloned into Escherichia coli and partially sequenced. This recombinant gene expresses a full-length 40-kDa product, which is recognized by a monoclonal antibody directed against the species-specific epitope of MOMP. The recombinant ompl is expressed in either insertion orientation, indicating that it utilizes its own promoter system. The endogenous ompl promoter possesses a relatively low activity despite the high level of MOMP expression. Deletion of a 520-bp fragment at the 3′ end encoding 39 amino acids (aa) at the C terminus and the remainder of the noncoding region leads to a significant decrease in mRNA stability and loss of protein synthesis. When the MOMP-encoding plasmid was introduced into E. coli minicells, it expressed 40- and 43-kDa proteins; however, inhibition of post-translational processing by ethanol revealed only a 43-kDa protein. These data indicate that the unprocessed ompl gene product contains a 22-aa leader sequence which is cleaved during translocation to the outer membrane, to yield a processed 40-kDa protein. The recombinant MOMP was localized to the outer membrane E. coli fraction, comparable to the location of the native C. trachomatis protein.

Original languageEnglish (US)
Pages (from-to)97-103
Number of pages7
JournalGene
Volume87
Issue number1
DOIs
StatePublished - Mar 1 1990
Externally publishedYes

Keywords

  • epitope
  • gene product
  • leader polypeptide
  • minicells
  • monoclonal antibodies
  • phage λ vectors
  • preprocessed polypeptide
  • Recombinant DNA

ASJC Scopus subject areas

  • Genetics

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