Bacterial surface protein expression

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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435 693, 435 697, 435177, 435209, 4352523, 514 6, 530350, 530395, 530400, 536 221, 536 231, 536 234, 536 237, 42419711, 4241921, C12P 2106, C07K 100, A61K 3800, C07H 1900

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055003532

ABSTRACT:
This invention provides a bacterium having an S-layer modified such that the bacterium S-layer protein gene contains one or more in-frame sequences coding for one or more heterologous polypeptides and, the S-layer is a fusion product of the S-layer protein and the heterologous polypeptide. The bacterium is preferably a Caulobacter which may be cultured as a film in a bioreactor or may be used to present an antigenic epitope to the environment of the bacterium. This invention also provides a method of expressing and presenting to the environment of a Caulobacter, a polypeptide that is heterologous to the S-layer of Caulobacter which comprises cloning a coding sequence for the polypeptide in-frame into an S-layer protein gene of Caulobacter whereby the polypeptide is expressed and presented on the surface of the Caulobacter as a fusion product of the S-layer protein and the polypeptide in the S-layer of the Caulobacter.

REFERENCES:
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Fisher, James A.; John Smit; and, Nina Agabian; "Transcriptional Analysis of the Major Surface Array Gene of Caulobacter crescentus" (1988) J. Bacteriol 170, 4706-4713.
Charbit, Alain; et al; "Versatility of a Vector for Expressing Foreign Polypeptides at the Surface of Gram-Negative Bacteria" (1988) Gene 70, 181-189.
Bingle, Wade H.; and, John Smit; "High-Level Expression Vectors for Caulobacter crescentus Incorporating the Transcription/Translation Initiation Regions of the Paracrystalline Surface-Layer-Protein Gene" (1990) Plasmid 24, 143-148.
Hayes, L. J.; et al; "Chlamydia trachomatis Major Outer Membrane Protein Epitopes Expressed as Fusions with LamB in an Attenuated aroA Strain of Salmonella typhimuirum; Their Application as Potential Immunogens" (1991) J. General Microbiology 137, 1557-1564.
Smit, John; and, James Atwater; "Use of Caulobacters to Separate Toxic Heavy Metals from Wastewater Streams" (Apr., 1991) U.S. Department of Energy publication.
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Bingle, W. H., et al, "Definition of Form and Function for the S-Layer of Caulobacter crescentus" in: Advances in Bacterial Paracrystalline Surface Layers (1993) Plenum Press (T. J. Beveridge and S. F. Koval ed.), pp. 181-197 (see enclosed author's copy insert at pp. 9-11).
Bingle, W. H., et al, "Linker Mutagenesis of the Caulobacter crescentus S-Layer Protein", in: Advances in Bacterial Paracrystalline Surface Layers (1993) Plenum Press (T. J. Beveridge and S. F. Koval ed.), pp. 293-296.
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