Vectors for constructing kan gene fusions: direct selection of mutations affecting IS10 gene expression.
Gene
; 90(1): 135-40, 1990 May 31.
Article
en En
| MEDLINE
| ID: mdl-2165970
ABSTRACT
We describe several vectors for constructing translational fusions to the kan gene of Tn5. Fusions are constructed in vitro using multi-copy vectors containing unique cloning sites situated between upstream transcriptional terminators and a downstream kan gene lacking transcriptional and translational start signals. Multi-copy fusions can be converted to single-copy chromosomal fusions by in vivo recombination with specific phage lambda vectors and vice versa. We find that kan fusions are often more suitable than lacZ fusions for the direct selection of mutations that increase fusion expression. These vectors were developed for isolating mutations that increase IS10 transposase expression; we describe strategies used to isolate such mutations, which map to IS10 or the Escherichia coli himA, himD(hip), dam or infC genes.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Elementos Transponibles de ADN
/
Kanamicina
/
Vectores Genéticos
Tipo de estudio:
Guideline
Idioma:
En
Revista:
Gene
Año:
1990
Tipo del documento:
Article