Construction of a novel kind of expression plasmid by homologous recombination in Saccharomyces cerevisiae.
Sci China C Life Sci
; 48(4): 330-6, 2005 Aug.
Article
em En
| MEDLINE
| ID: mdl-16248426
ABSTRACT
Based on a previously used plasmid pHC11, a new plasmid pHC11R was constructed. Cutting plasmid pHC11R with proper restriction enzymes, the resulting larger DNA fragment pHC11R' was co-transformed with a PCR amplified expression cassette of human IFNalpha2b into yeast. By means of the homologous sequences at both ends of two DNA fragments, a novel expression plasmid pHC11R-IFNalpha2b was formed via homologous recombination in the yeast. Compared with pHC11-IFNalpha2b, the expression plasmid pHC11R-IFNalpha2b was smaller in size and in absence of antibiotic resistant gene. The stability and copy number of pHC11R-IFNalpha2b were greatly increased and the expression level of heterologous protein was improved. As the derivatives of pHC11R, a series of recombination expression vectors pHRs containing different combination of expression elements were developed. This led to a rapid and powerful method for cloning and expressing of different genes in yeast.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Plasmídeos
/
Recombinação Genética
/
Saccharomyces cerevisiae
/
Vetores Genéticos
Limite:
Humans
Idioma:
En
Revista:
Sci China C Life Sci
Assunto da revista:
BIOLOGIA
/
CIENCIA
Ano de publicação:
2005
Tipo de documento:
Article
País de afiliação:
China