Gene knockout and knockin on the Escherichia coli lac operon loci using pBR322-red system / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 192-197, 2005.
Article
Dans Chinois
| WPRIM
| ID: wpr-249926
ABSTRACT
pBR322-Red is a newly constructed recombineering plasmid, which contains a part of the pBR322 vector, a series of regulatory elements of lambda-prophage and Red recombination genes. In the beginning, we studied the best working conditions of pBR322-Red, and then modified lac operon in E. coli W3110 chromosome using the plasmid as follow Firstly, we knockout the lacI gene using Red-mediated recombineering with overlapping single stranded DNA oligonucleotides. Secondly, we substituded the lacA and lacY genes with lacZ, a report gene, by Red-mediated linearized double strands DNA homologous recombination. Finally, we detected the expression of lacZ on these loci for the first time. The results suggested that pBR322-Red system is suitable for modifying W3110 chromosome with various recombination strategies.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Plasmides
/
Recombinaison génétique
/
Chromosomes de bactérie
/
Bactériophage lambda
/
Escherichia coli
/
Techniques de knock-in de gènes
/
Techniques de knock-out de gènes
/
Génétique
/
Opéron lac
/
Méthodes
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Chinese Journal of Biotechnology
Année:
2005
Type:
Article
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