High-Yield Expression in Escherichia coli and Purification of Mouse Ubiquitin-Activating Enzyme E1

• Tipo: Artigo em Revista Científica Internacional
• Data: 2012

Autores

carvalho, af (Autor) Outra; pinto, mp (Autor) Outra; grou, cp (Autor) Outra; vitorino, r (Autor) Outra; domingues, p (Autor) Outra; yamao, f (Autor) Outra; sá-miranda, c (Autor) Outra; azevedo, je (Autor) FCUP

Revista

Título: Molecular Biotechnology

Vol. 51

Páginas: 1-8

ISSN: 1073-6085

Editora: Springer Nature

Indexação

ISI Web of Knowledge - 3 Citações

Scopus - 3 Citações

Outras Informações

ID Authenticus: P-002-8RY

DOI: 10.1007/s12033-011-9463-x

Abstract (EN): Research in the ubiquitin field requires large amounts of ubiquitin-activating enzyme (E1) for in vitro ubiquitination assays. Typically, the mammalian enzyme is either isolated from natural sources or produced recombinantly using baculovirus/insect cell protein expression systems. Escherichia coli is seldom used to produce mammalian E1 probably due to the instability and insolubility of this high-molecular mass protein. In this report, we show that 5-10 mg of histidine-tagged mouse E1 can be easily obtained from a 1 l E. coli culture. A low temperature during the protein induction step was found to be critical to obtain an active enzyme.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 8