Abstract
A HindII restriction fragment comprising the Escherichia coli lac regulatory region and the genetic information for the alpha peptide of beta-galactosidase (beta-D-galactosidegalactohydrolase, EC. 3.2.1.23) has been inserted into 1 of the 10 Bsu I cleavage sites of M13 by blunt end ligation. A stable hybrid phage was isolated and identified by its ability to complement the lac alpha function. Further characterization of the hybrid phage includes retransformation studies, agarose gel electrophoresis, DNA-DNA hybridization, and heteroduplex mapping. The insertion point has been localized at 0.083 map unit on thewild-type circular map-i.e., within the intergenic region. The results prove that part of the intergenic region is nonessential and that the phage can be used as a cloning vehicle.
Keywords
HeteroduplexBiologyProphageMolecular biologyGeneticsRestriction enzymeDNAIntergenic regionAgarose gel electrophoresisEscherichia coliGeneBacteriophageGenome
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Publication Info
- Year
- 1977
- Type
- article
- Volume
- 74
- Issue
- 9
- Pages
- 3642-3646
- Citations
- 707
- Access
- Closed
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Cite This
Joachim Messing,
Bruno Gronenborn,
Benno Müller‐Hill
et al.
(1977).
Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro..
Proceedings of the National Academy of Sciences
, 74
(9)
, 3642-3646.
https://doi.org/10.1073/pnas.74.9.3642
Identifiers
- DOI
- 10.1073/pnas.74.9.3642