A cloning vector employing a versatile ß-glucosidase as an indicator for recombinant clones.
Anal Biochem
; 425(2): 166-8, 2012 Jun 15.
Article
em En
| MEDLINE
| ID: mdl-22425541
ABSTRACT
A mutant glucosidase, cpGluT, with activity toward chromogenic substrates (X-gal [5-bromo-4-chloro-3-idolyl-ß-d-galactoside] and indican) and a fluorogenic 4-methylumbeliferyl-ß-d-glucopyranoside (MUG) was constructed by replacing the monomeric ß-glucosidase region (E314-N326) with designed multiple cloning sites. When expressed in hosts (lacZ+ and lacZ-), a vector containing the cpGluT produced a colored or fluorescent phenotype according to the substrate supplemented on LB plates without any inducer. cpGluT is readily incorporable into customized vectors and does not require special hosts to detect recombinant plasmids, thereby making screening recombinants more effective and less expensive.
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Galactosídeos
/
Vetores Genéticos
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
2012
Tipo de documento:
Article