A Novel Chimeric MOMP Antigen Expressed in Escherichia Coli, Arabidopsis Thaliana, and Daucus Carota as a Potential Chlamydia Trachomatis Vaccine Candidate

Protein Expr Purif. 2011 Dec;80(2):194-202. doi: 10.1016/j.pep.2011.08.010. Epub 2011 Aug 28.

Abstract

The major outer membrane protein (MOMP) of Chlamydia trachomatis is a highly antigenic and hydrophobic transmembrane protein. Our attempts to express the full-length protein in a soluble form in Escherichia coli and in transgenic plants failed. A chimeric gene construct of C. trachomatis serovar E MOMP was designed in order to increase solubility of the MOMP protein but with retained antigenicity. The designed construct was successfully expressed in E. coli, in Arabidopsis thaliana, and in Daucus carota. The chimeric MOMP expressed in and purified from E. coli was used as antigen for production of antibodies in rabbits. The anti-chimeric MOMP antibodies recognized the corresponding protein in both E. coli and in transgenic plants, as well as in inactivated C. trachomatis elementary bodies. Transgenic Arabidopsis and carrots were characterized for the number of MOMP chimeric genetic inserts and for protein expression. Stable integration of the transgene and the corresponding protein expression were demonstrated in Arabidopsis plants over at least six generations. Transgenic carrots showed a high level of expression of the chimeric MOMP - up to 3% of TSP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium / genetics
  • Agrobacterium / metabolism
  • Amino Acid Sequence
  • Animals
  • Antigen-Antibody Complex / immunology
  • Antigens, Bacterial / genetics
  • Antigens, Bacterial / immunology*
  • Antigens, Bacterial / isolation & purification
  • Antigens, Bacterial / metabolism
  • Arabidopsis / genetics
  • Arabidopsis / metabolism
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / immunology*
  • Bacterial Outer Membrane Proteins / isolation & purification
  • Bacterial Outer Membrane Proteins / metabolism
  • Chlamydia trachomatis / genetics
  • Chlamydia trachomatis / immunology*
  • Chlamydia trachomatis / metabolism
  • Cloning, Molecular
  • Daucus carota / genetics
  • Daucus carota / metabolism
  • Electroporation
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Genome, Bacterial
  • Immunoblotting
  • Molecular Sequence Data
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Rabbits
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Solubility
  • Transformation, Genetic
  • Transgenes

Substances

  • Antigen-Antibody Complex
  • Antigens, Bacterial
  • Bacterial Outer Membrane Proteins
  • Recombinant Proteins
  • OMPA outer membrane proteins