A general method to select for M13 clones carrying base pair substitution mutants constructedin vitro
- 1 January 1983
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 11 (12) , 4229-4239
- https://doi.org/10.1093/nar/11.12.4229
Abstract
In this paper we describe a method to select base pair substitution mutants constructed in vitro. The mutagenesis is performed by forcing mistakes during in vitro synthesis from a primer annealed to a single stranded DNA template. The selection is based on the fact that, following transformation, the progeny of the strand elongated in vitro and the template strand have different phenotypes. The method is general and applicable to any DNA segment; the type of base pair substitution and its position can be chosen at will. The combined efficiency of mutagenesis and selection allows for 85% frequency of mutants in all analyzed clones.Keywords
This publication has 13 references indexed in Scilit:
- Relationship between the two components of the split promoter of eukaryotic tRNA genes.Proceedings of the National Academy of Sciences, 1982
- Gap misrepair mutagenesis: efficient site-directed induction of transition, transversion, and frameshift mutations in vitro.Proceedings of the National Academy of Sciences, 1982
- Site-directed mutagenesis of a tRNA gene: base alterations in the coding region affect transcription.Proceedings of the National Academy of Sciences, 1982
- Site-specific mutagenesis by error-directed DNA synthesisNature, 1982
- Directed mutagenesis of DNA cloned in filamentous phage: influence of hemimethylated GATC sites on marker recovery from restriction fragmentsNucleic Acids Research, 1982
- Transcription of tRNA genes in vivo: single-stranded compared to double-stranded templates.Proceedings of the National Academy of Sciences, 1980
- Cloning of nematode tRNA genes and their expression in the frog oocyte.1978
- DNA sequencing with chain-terminating inhibitorsProceedings of the National Academy of Sciences, 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, 1977
- Induction and isolation of mutants in a specific region of gene A of bacteriophage ∅X174Virology, 1976