pUR222, a vector for cloning and rapid chemical sequencing of DNA
Open Access
- 25 August 1981
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 9 (16) , 4087-4098
- https://doi.org/10.1093/nar/9.16.4087
Abstract
A multipurpose plasmid, pUR222, was constructed. It contains six unique cloning sites (PstI, SalI, AccI, HindII, BamHI and EcoRI) in a small region of its lacZ-gene part. Insertion of foreign DNA into the plasmid can be easily detected. Bacteria harbouring recombinant plasmids generally give rise to w hite colonies, while those containing only vector DNA form blue colonies on indicator plates. Plasmid DNA purified by a rapid method (Birnboim, H. C. and Doly, J. (1979) Nucl. Acids. Res. 7, 1513–1523) can be used for chemical sequencing of the cloned insert DNA. Labeled fragments need not be isolated after cutting with the proper restriction enzymes and are treated directlyaccording to the sequencing protocol of Maxam and Gilbert.Keywords
This publication has 19 references indexed in Scilit:
- Complete Nucleotide Sequence of the Escherichia coli Plasmid pBR322Published by Cold Spring Harbor Laboratory ,1979
- The insertion element IS1 is a natural constituent of coliphage P1 DNAPlasmid, 1978
- pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs longNucleic Acids Research, 1978
- DNA sequencing with chain-terminating inhibitorsProceedings of the National Academy of Sciences, 1977
- Construction and characterization of new cloning vehicle. II. A multipurpose cloning systemGene, 1977
- Construction and characterization of new cloning vehicles I. Ampicillin-resistant derivatives of the plasmid pMB9Gene, 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
- A rapid method for desalting small volumes of solutionAnalytical Biochemistry, 1973
- Calcium-dependent bacteriophage DNA infectionJournal of Molecular Biology, 1970
- Growth Direction and Segregation of the E. coli ChromosomeCold Spring Harbor Symposia on Quantitative Biology, 1968