Cloning, sequencing, and expression of the gene coding for an antigenic 120-kilodalton protein of Rickettsia conorii

K. W. Schuenke, David Walker

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Several high-molecular-mass (above 100 kDa) antigens are recognized by sera from humans infected with spotted fever group rickettsiae and may be important stimulators of the host immune response. Molecular cloning techniques were used to make genomic Rickettsia conorii (Malish 7 strain) libraries in expression vector λgt11. The 120-kDa R. conorii antigen was identified by monospecific antibodies to the recombinant protein expressed on construct λ4-7. The entire gene DNA sequence was obtained by using this construct and two other overlapping constructs. An open reading frame of 3,068 bp with a calculated molecular mass of approximately 112 kDa was identified. Promoters and a ribosome-binding site were identified on the basis of their DNA sequence homology to other rickettsial genes and their relative positions in the sequence. The DNA coding region shares no significant homology with other spotted fever group rickettsial antigen genes (i.e., the R. rickettsii 190-, 135-, and 17-kDa antigen-encoding genes). The PCR technique was used to amplify the gene from eight species of spotted fever group rickettsiae. A 75-kDa portion of the 120-kDa antigen was overexpressed in and purified from Escherichia coli. This polypeptide was recognized by antirickettsial antibodies and may be a useful diagnostic reagent for spotted fever group rickettsioses.

Original languageEnglish (US)
Pages (from-to)904-909
Number of pages6
JournalInfection and Immunity
Volume62
Issue number3
StatePublished - 1994
Externally publishedYes

Fingerprint

Rickettsia conorii
Organism Cloning
Gene Expression
Antigens
Fever
Rickettsia
Proteins
Genes
vpr Genes
Antibodies
Molecular Cloning
Sequence Homology
Ribosomes
Recombinant Proteins
Open Reading Frames
Libraries
Binding Sites
Escherichia coli
Polymerase Chain Reaction
Peptides

ASJC Scopus subject areas

  • Immunology

Cite this

Cloning, sequencing, and expression of the gene coding for an antigenic 120-kilodalton protein of Rickettsia conorii. / Schuenke, K. W.; Walker, David.

In: Infection and Immunity, Vol. 62, No. 3, 1994, p. 904-909.

Research output: Contribution to journalArticle

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