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Nucleic Acids Research, 1975, Vol. 2, No. 12 2365-2378
© 1975

Articles

Insertion of a rabbit ß-globin gene sequence into an E.coli plasmid

F. Rougeon, P. Kourilsky* and B. Mach

Department of Pathology, University of Geneva Geneva, Switzerland

Received November 10, 1975.

Double stranded DNA has been synthesized in vitro from rabbit globin messenger RNA and elongated with homopolymeric dG tails. An E.coli plasmid was cleaved by EcoRI. The cohesive ends were repaired and dC tails added, to permit reconstitution of the EcoRI sites upon annealing with the dG elongated globin DNA. Transformation of E.coli with the globin-plasmid DNA hybrid has yielded a clone which harbours a recombinant plasmid (pCR1-BG1), as demonstrated by hybridization experiments with radioactive globin cDNA. The sequence carried by the recombinant plasmid corresponds to part of the gene sequence coding for the ß chain of rabbit globin. Circular DNA of the purified recombinant plasmid exhibits sensitivity to EcoRI.

*Department of Molecular Biology, Pasteur Institute, Paris, France


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