pUR222, a vector for cloning and rapid chemical sequencing of DNA.
AUTOR(ES)
Rüther, U
RESUMO
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 lac Z-gene part. Insertion of foreign DNA into the plasmid can be easily detected. Bacteria harbouring recombinant plasmids generally give rise to white 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 directly according to the sequencing protocol of Maxam and Gilbert.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=327416Documentos Relacionados
- pUR 250 allows rapid chemical sequencing of both DNA strands of its inserts.
- pSEMCatR1: a procaryotic-eucaryotic shuttle vector compatible with pUR and lambda gt11 expression systems.
- pDUAL: a transposon-based cosmid cloning vector for generating nested deletions and DNA sequencing templates in vivo.
- Non-radioactive chemical sequencing of biotin labelled DNA.
- Pyrrolidine, a non-controlled substance, can replace piperidine for the chemical sequencing of DNA.