A new idea for simple and rapid monitoring of gene expression: requirement of nucleotide sequences encoding an N-terminal HA tag in the T7 promoter-driven expression in E. coli.
Biotechnol Lett
; 34(10): 1841-6, 2012 Oct.
Article
em En
| MEDLINE
| ID: mdl-22714269
ABSTRACT
Mammalian expression vectors are used to overexpress genes of interest in mammalian cells. High temperature requirement protein A1 (HtrA1), used as a specific target, was expressed from the pHA-M-HtrA1 plasmid in HEK293T cells, inducing cell death. Expression of HtrA1 was driven by the pHA-M-HtrA1 mammalian expression vector in E. coli resulting in growth suppression of E. coli in an HtrA1 serine protease-dependent manner. By using various combinations of promoters, target genes and N-terminal tags, the T7 promoter and N-terminal HA tag in the mammalian expression vector were shown to be responsible for expression of target genes in E. coli. Thus the pHA-M-HtrA1 plasmid can be used as a novel, rapid pre-test system for expression and cytotoxicity of the specific target gene in E. coli before assessing its functions in mammalian cells.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Biotecnologia
/
Proteínas Recombinantes de Fusão
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Serina Endopeptidases
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Glicoproteínas de Hemaglutininação de Vírus da Influenza
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Escherichia coli
Limite:
Humans
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article