Applied Microbiology and Biotechnology

, Volume 25, Issue 3, pp 267–271 | Cite as

A new expression vector for the production of fused proteins in Escherichia coli

  • Noemi Flores
  • Ramon de Anda
  • Leopoldo Guereca
  • Norberto Cruz
  • Salvador Antonio
  • Paulina Balbas
  • Francisco Bolivar
  • Fernando Valle
Article

Summary

The construction of a new expression vector for fused proteins production in Escherichia coli is reported. This new vehicle uses the trp promoter-operator control region for the high level expression of a DNA fragment that codes for the amino terminal fragment of the cI λ repressor protein. This truncated polypeptide is accumulated as inclusion bodies that are easily purified. To probe the benefits of the system, synthetic DNA that codes for the human insulin B chain, was cloned at the end of the DNA coding region for the cI truncated peptide. The hybrid peptide thus produced after induction, allowed an easy and reproducible purification of active insulin B chain.

Abbreviation

Ap

ampicillin

bp

base pairs

kD

kilodaltons

Mr

migration rate

PAGE

polyacrylamide gel electrophoresis

Tc

tetracycline

trp

tryptophan

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References

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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Noemi Flores
    • 1
  • Ramon de Anda
    • 1
  • Leopoldo Guereca
    • 1
  • Norberto Cruz
    • 1
  • Salvador Antonio
    • 1
  • Paulina Balbas
    • 1
  • Francisco Bolivar
    • 1
  • Fernando Valle
    • 1
  1. 1.Departamentos de Biologia Molecular y Bioquimica, Centro de Investigaciones sobre Ingenieria Genetica y BiotecnologiaUNAMCuernavaca MorelosMexico

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