A general method for cloning eukaryotic structural gene sequences.
- 1 September 1976
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 73 (9) , 3146-3150
- https://doi.org/10.1073/pnas.73.9.3146
Abstract
Complementary DNA, transcribed in vitro from purified rabbit globin mRNA and made double-stranded, was inserted into Escherichia coli plasmids pSC101 and pMB9 by the poly(dT)/poly(dA) tailing and annealing technique. E. coli transformants given by this DNA preparation contained globin sequences as shown by the hybridization of globin RNA to DNA from clones grown and lysed in situ on nitrocellulose filters. An estimate of the amount of inserted globin sequences was provided by fingerprint analysis of globin mRNA sequences hybridized to the purified plasmid chimeras. Inserted sequences so far subjected to detailed analysis were ascribed to the rabbit .beta. globin chain. The susceptibility of inserted .beta. globin sequences to the EcoRI restriction endonuclease confirms the existence of a site already found through previous nucleotide sequence analysis.Keywords
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