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Molecular Pathogenesis

Temperature-Regulated Protein Synthesis by Leptospira interrogans

Jarlath E. Nally, John F. Timoney, Brian Stevenson
Jarlath E. Nally
Department of Veterinary Medicine, University of Kentucky, Lexington, Kentucky 40546-0099,and
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John F. Timoney
Department of Veterinary Medicine, University of Kentucky, Lexington, Kentucky 40546-0099,and
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Brian Stevenson
Department of Microbiology and Immunology, University of Kentucky, Lexington, Kentucky 40536-0298
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DOI: 10.1128/IAI.69.1.400-404.2001
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    Fig. 1.

    (A) Antigens in lysates of L. interrogansserovar pomona type kennewicki JEN4 grown at 30°C and shifted to 24, 30, or 37°C. The lysates were separated by SDS-PAGE and immunoblotted with serum from a convalescent mare. A 57-kDa antigen apparently upregulated at 37°C is indicated with an arrow. The asterisks indicate antigens present in greater amounts at 24 and 30°C than at 37°C. (B) To establish that equivalent amounts of protein were loaded in the gel shown in panel A, lysates were probed with antiserum to endoflagella of T. pallidum, a non-temperature-regulated protein. Molecular mass markers are indicated in kilodaltons.

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    Fig. 2.

    Immunoblot analysis of leptospiral heat shock proteins DnaK and GroEL in lysates of L. interrogans JEN4 expressed at 30 and 37°C following SDS-PAGE. Molecular mass markers are indicated in kilodaltons.

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    Fig. 3.

    (A) Silver stain of Triton X-114 extracts of L. interrogans JEN4 at 30 or 37°C separated by SDS-PAGE and showing proteins in fractions consisting of protoplasmic cylinder (P), aqueous phase (A), and detergent phase (D). (B) Fractions probed with antiserum to T. pallidum endoflagella to demonstrate that the protoplasmic cylinder was intact. Molecular mass markers are indicated in kilodaltons. Upregulated proteins are indicated with arrows; downregulated proteins are indicated with asterisks.

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    Fig. 4.

    Triton X-114 extract of L. interrogans JEN4 at 30 and 37°C immunoblotted with serum from a convalescent mare following SDS-PAGE. Fractions consisting of protoplasmic cylinder (P), aqueous phase (A), and detergent phase (D) are reactive. Molecular mass markers are indicated in kilodaltons. Upregulated antigens are indicated with arrows; downregulated antigens are indicated with asterisks.

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    Fig. 5.

    Lysates of JEN4 grown at 30°C, grown at 30°C and shifted to 37°C, grown at 37°C and maintained at 37°C, or grown at 37°C and shifted to 30°C. Following SDS-PAGE, the lysates were immunoblotted with LipL36- and LipL41-specific antisera.

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Temperature-Regulated Protein Synthesis by Leptospira interrogans
Jarlath E. Nally, John F. Timoney, Brian Stevenson
Infection and Immunity Jan 2001, 69 (1) 400-404; DOI: 10.1128/IAI.69.1.400-404.2001

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Temperature-Regulated Protein Synthesis by Leptospira interrogans
Jarlath E. Nally, John F. Timoney, Brian Stevenson
Infection and Immunity Jan 2001, 69 (1) 400-404; DOI: 10.1128/IAI.69.1.400-404.2001
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KEYWORDS

Antigens, Bacterial
Bacterial Proteins
Leptospira interrogans

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