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.
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Abbreviations
- Ap:
-
ampicillin
- bp:
-
base pairs
- kD:
-
kilodaltons
- Mr:
-
migration rate
- PAGE:
-
polyacrylamide gel electrophoresis
- Tc:
-
tetracycline
- trp:
-
tryptophan
References
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Flores, N., de Anda, R., Guereca, L. et al. A new expression vector for the production of fused proteins in Escherichia coli . Appl Microbiol Biotechnol 25, 267–271 (1986). https://doi.org/10.1007/BF00253661
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DOI: https://doi.org/10.1007/BF00253661