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Nucleic Acids Research, 1978, Vol. 5, No. 12 4479-4494
© 1978

Articles

Bacteriophage lambda and plasmid vectors, allowing fusion of cloned genes in each of the three translational phases

Patrick Charnay*, Michel Perricaudet*, Francis Galibert** and Pierre Tiollais*

*Recombinaison et expression génétique (INSERM U. 163) Unité de Génie Génétique, Institut Pasteur 28, rue du Docteur Roux, 75015 Paris **Laboratoire d'Hématologie Expérimentale, Centre HAYEM, Hôpital Saint-Louis 2, place du Dr. A. Fournier 75010 Paris, France

Received November 13, 1978. We have constructed vectors from bacteriophage lambda and from plasmid pBR322 having a single EcoRI restriction site which is immediatly downstream from the lac UV5 promotor. Each vector allows the fusion of a cloned gene to the lac Z gene in a different phase relative to the translation initiation codon of the lac Z gene. These vectors were constructed through modification of the initial EcoRI restriction site by S1 endonuclease treatment and then addition of octadeoxyribonucleotides (EcoRI linkers) , which shifted the restriction site by 2 or 4 nucleotides. Used in combination these vectors should allow translation of a cloned gene in any one of the three coding phases. The bacteriophages vectors are certified as B2 (EK2) safety level vectors by the French "recombinaison génétique in vitro" committee (D.G.R.S.T.).


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