Insertion of a rabbit β-globin gene sequence into an E.coli plasmid
- 1 January 1975
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 2 (12) , 2365-2378
- https://doi.org/10.1093/nar/2.12.2365
Abstract
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-betaG1), 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 beta chain of rabbit globin. Circular DNA of the purified recombinant plasmid exhibits sensitivity to EcoRI.Keywords
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