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Simple piggyBac transposon-based mammalian cell expression system for inducible protein production.
Li, Zhijie; Michael, Iacovos P; Zhou, Dongxia; Nagy, Andras; Rini, James M.
Afiliação
  • Li Z; Department of Biochemistry, University of Toronto, Toronto, ON, Canada M5S 1A8.
Proc Natl Acad Sci U S A ; 110(13): 5004-9, 2013 Mar 26.
Article em En | MEDLINE | ID: mdl-23476064
ABSTRACT
Reported here is a piggyBac transposon-based expression system for the generation of doxycycline-inducible, stably transfected mammalian cell cultures for large-scale protein production. The system works with commonly used adherent and suspension-adapted mammalian cell lines and requires only a single transfection step. Moreover, the high uniform expression levels observed among clones allow for the use of stable bulk cell cultures, thereby eliminating time-consuming cloning steps. Under continuous doxycycline induction, protein expression levels have been shown to be stable for at least 2 mo in the absence of drug selection. The high efficiency of the system also allows for the generation of stable bulk cell cultures in 96-well format, a capability leading to the possibility of generating stable cell cultures for entire families of membrane or secreted proteins. Finally, we demonstrate the utility of the system through the large-scale production (140-750 mg scale) of an endoplasmic reticulum-resident fucosyltransferase and two potential anticancer protein therapeutic agents.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes / Elementos de DNA Transponíveis / Expressão Gênica / Elementos de Resposta Limite: Animals / Humans Idioma: En Revista: Proc Natl Acad Sci U S A Ano de publicação: 2013 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Recombinantes / Elementos de DNA Transponíveis / Expressão Gênica / Elementos de Resposta Limite: Animals / Humans Idioma: En Revista: Proc Natl Acad Sci U S A Ano de publicação: 2013 Tipo de documento: Article