Using recombineering to generate point mutations:galK-based positive-negative selection method.
Methods Mol Biol
; 852: 121-31, 2012.
Article
em En
| MEDLINE
| ID: mdl-22328430
ABSTRACT
Recombineering is a recombination-based highly efficient method of genetic engineering. It can be used to manipulate the bacterial chromosomal DNA as well as any episomal DNA. Recombineering can be used to insert selectable or nonselectable DNA fragments and subclone DNA fragments without the use of restriction enzymes and also to make precise alterations including single nucleotide changes in the DNA. Here we describe a galactokinase (galK)-based two-step method to generate point mutations in the bacterial artificial chromosome (BAC) insert using the recombineering technology. It takes advantage of the ability to select and also counterselect for the presence of galK.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Seleção Genética
/
DNA Recombinante
/
Engenharia Genética
/
Mutagênese
/
Mutação Puntual
/
Galactoquinase
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Estados Unidos