Opening book details…
Can I read Purification and characterization of pea thioredoxin f expressed in Escherichia coli on EtoBox?
Purification and characterization of pea thioredoxin f expressed in Escherichia coli by Michael Hodges; Myroslawa Miginiac-Maslow; Paulette Decottignies; Jean-Pierre Jacquot; Mariana Stein; Loic Lepiniec; Claude Crétin; Pierre Gadal is a Agricultural and Biological Sciences article available to read on EtoBox.
What is Purification and characterization of pea thioredoxin f expressed in Escherichia coli about?
The recently cloned cDNA for pea chloroplast thioredoxin f was used to produce, by PCR, a fragment coding for a protein lacking the transit peptide. This cDNA fragment was subcloned into a pET expression vector and used to transform E. coli cells. After induction with IPTG the transformed cells produce the protein, mainly in the soluble fraction of the broken cells. The recombinant thioredoxin f has been purified and used to raise antibodies and analysed for activity. The antibodies appear to be specific towards thioredoxin f and do not recognize other types of thioredoxin. The recombinant protein could activate two chloroplastic enzymes, namely NADP-dependent malate dehydrogenase (NADP-MDH) and fructose 1,6-bisphosphatase (FBPase), both using dithiothreitol as a chemical reductant and in a light-reconstituted/thylakoid assay. Recombinant pea thioredoxin f turned out to be an excellent catalyst for NADP-MDH activation, being the more efficient than a recombinant m-type thioredoxin of Chlamydomonas reinhardtii and the thioredoxin of E. coli. At the concentrations of thioredoxin used in the target enzyme activation assays only the recombinant thioredoxin f activated the FBPase.
Who reads Purification and characterization of pea thioredoxin f expressed in Escherichia coli?
It is typically read by researchers, students, and practitioners in Agricultural and Biological Sciences.
- Author
- Michael Hodges; Myroslawa Miginiac-Maslow; Paulette Decottignies; Jean-Pierre Jacquot; Mariana Stein; Loic Lepiniec; Claude Crétin; Pierre Gadal
- Publisher
- Springer; Springer-Verlag; Kluwer Academic Publishers; Springer Science and Business Media LLC; Society for Mining, Metallurgy and Exploration Inc. (ISSN 0167-4412)
- Published
- 1994
- Language
- EN
- Field
- Agricultural and Biological Sciences (Life Sciences)