Rapid and efficient cloning of proviral flanking fragments by kanamycin resistance gene complementation.
Nucleic Acids Res
; 27(2): 706-7, 1999 Jan 15.
Article
en En
| MEDLINE
| ID: mdl-9863001
ABSTRACT
We have developed a technique for the rapid cloning of unknown flanking regions of transgenic DNA. We complemented a truncated kanamycin resistance gene of a bacterial plasmid with a neomycin resistance gene fragment from a gene transfer vector. Optimized transformation conditions allowed us to directly select for kanamycin-resistant bacteria. We cloned numerous proviral flanking fragments from growth factor-independent cell mutants that were obtained after infection with a replication incompetent retroviral vector and identified integrations into the cyclin D2 and several unknown genomic sequences. We anticipate that our method could be adapted to various vector systems that are used to tag and identify genes and to map genomes.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Retroviridae
/
Resistencia a la Kanamicina
/
Provirus
/
Clonación Molecular
/
Prueba de Complementación Genética
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
Nucleic Acids Res
Año:
1999
Tipo del documento:
Article
País de afiliación:
Alemania