Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro.
- 1 September 1977
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 74 (9), 3642-3646
- https://doi.org/10.1073/pnas.74.9.3642
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.This publication has 23 references indexed in Scilit:
- Construction of plasmids carrying the cI gene of bacteriophage lambda.Proceedings of the National Academy of Sciences of the United States of America, 1976
- Mapping of Promoter Sites on the Genome of Bacteriophage M13JBIC Journal of Biological Inorganic Chemistry, 1976
- Synthetic lacoperator DNA is functional in vivoNature, 1976
- Restriction-Enzyme-Cleavage Maps of Bactenophage M13. Existence of an Intergenic Region on the M13 GenomeJBIC Journal of Biological Inorganic Chemistry, 1976
- Overproduction of phage Lambda repressor under control of the lac promotor of Escherichia coliMolecular Genetics and Genomics, 1976
- Viable Molecular Hybrids of Bacteriophage Lambda and Eukaryotic DNAProceedings of the National Academy of Sciences of the United States of America, 1974
- Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNAProceedings of the National Academy of Sciences of the United States of America, 1974
- Replication and Transcription of Eukaryotic DNA in Esherichia coliProceedings of the National Academy of Sciences of the United States of America, 1974
- Biochemical Method for Inserting New Genetic Information into DNA of Simian Virus 40: Circular SV40 DNA Molecules Containing Lambda Phage Genes and the Galactose Operon of Escherichia coliProceedings of the National Academy of Sciences of the United States of America, 1972
- Inhibition of Minicircular DNA Replication in Escherichia coli 15 by RifampicinNature New Biology, 1972