A plasmid expression vector that permits stabilization of both mRNAs and proteins encoded by the cloned genes

Robert M. Duvoisin, Dominique Belin, Henry M. Krisch

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Two new expression vectors have been constructed to take advantage of several useful properties of bacteriophage T4-infected Escherichia coli.These plasmids,pRDB8 and pRDB9,contain the promoter region and start codon of T4 gene 32, a contiguous multiple cloning site (MCS),and translation and transcription termination signals. DNA fragments inserted into the MCS are transcribed and translated at a high level in both uninfected and phage T4-infected cells.Furthermore, the extreme stability of the hybrid mRNA after infection permits the specific biosynthetic labeling of the protein encoded by the cloned gene.In addition, the cloned gene product is stabilized, since the host-mediated degradation of foreign proteins is inhibited by phage infection. The properties of this expression system were demonstrated with the constant region of a rabbit immunoglobulin λ light chain (Cλ) gene.Although proteolytic degradation of the Cλfusion protein was rapid in uninfected cells,degradation was blocked in phage-infected cells and the protein accumulated in greater amounts.

Original languageEnglish (US)
Pages (from-to)193-201
Number of pages9
JournalGene
Volume45
Issue number2
DOIs
StatePublished - 1986

Keywords

  • Recombinant DNA
  • bacteriophage T4 gene 32
  • immunoglobulin lambda chain
  • protease inhibition

ASJC Scopus subject areas

  • Genetics

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