title: Use of lactose to induce expression of soluble NifA protein domains of Herbaspirillum seropedicae in Escherichia coli.
abstract: Overexpression and purification are procedures used to allow functional and structural characterization of proteins. Many overexpressed proteins are partially or completely insoluble, and can not be easily purified. The NifA protein is an enhancer-binding protein involved in activating the expression of nif and some fix genes. The NifA protein from many organisms is usually insoluble when over-expressed, and therefore difficult to work with in vitro. In this work we have overexpressed the central + C-terminal and the central domains of the Herbaspirrilum seropedicae NifA protein in an Escherichia coli background. Expression was induced with either IPTG or lactose. The data showed that induction with lactose promoted a significantly higher percentage of these proteins in the soluble fraction than with IPTG. This probably reflects a slower kinetics of induction by lactose.
authors: Department of Biochemistry, Universidade Federal do Paraná, Curitiba, Brazil.
authors: R A
authors: RA
authors: Monteiro
authors: E M
authors: EM
authors: Souza
authors: M G
authors: MG
authors: Yates
authors: F O
authors: FO
authors: Pedrosa
authors: L S
authors: LS
authors: Chubatsu
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D020562:Betaproteobacteria / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007785:Lactose / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C061338:NifA protein, Bacteria; D011994:Recombinant Proteins; D014157:Transcription Factors; D007785:Lactose