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Secretion Cloning Vectors for Guiding the Localization of Proteins in Vivo

  • C. A. Lunn
  • M. Takahara
  • M. Inouye
Part of the Current Topics in Microbiology and Immunology book series (CT MICROBIOLOGY, volume 125)

Abstract

Cloning of genes into high copy number vectors has supplied biochemists with a powerful method for the production and purification of large amounts of biochemically important macromolecules. Judicious choice of promoter sequences has produced vectors capable of loading cells with 50% of the total cellular protein as the desired gene product, making purification a trivial chore. However, this approach is limited to proteins whose activity is not detrimental to protein production, and to proteins not sensitive to cytoplasmic proteases (see below). Recognition of this problem has stimulated the development of several cloning vectors designed to guide a cloned gene product out of the cytoplasm of the bacterial cell. These vectors produce a hybrid protein in which the cloned gene product is fused with a signal peptide, a structural element necessary for protein translocation. In this review, we will describe the advantages and limitations of some currently available secretion cloning vectors, and will discuss the use of these tools to produce large amounts of protein and to determine the information required to direct proteins to have various compartments of the cell.

Keywords

Outer Membrane Cytoplasmic Membrane Outer Membrane Protein Total Cellular Protein Hybrid Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Berlin · Heidelberg 1986

Authors and Affiliations

  • C. A. Lunn
    • 1
  • M. Takahara
    • 2
  • M. Inouye
    • 1
  1. 1.Department of BiochemistryState University of New YorkStony BrookUSA
  2. 2.Biochemical Research LaboratoryTokyo Jozo Co. Ltd.Tagata-Gun, Shizuoka-KenJapan

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