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A ferredoxin-type iron-sulfur protein gene,frx B,is expressed in the chloroplasts of tobacco and spinach
Authors:Cheng-Hui Lin  Madeline Wu
Affiliation:(1) Institute of Molecular Biology, Academia Sinica, Nankang, 11529 Taipei, Republic of China;(2) Department of Biological Sciences, University of Maryland, Baltimore County Campus, 21228 Baltimore, MD, USA;(3) Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 21202 Baltimore, MD, USA
Abstract:Previously, a ferredoxin-type iron-sulfur protein, frx B protein, was identified in a high-salt extract of the purified thylakoid membrane of Chlamydomonas reinhardtii, a unicellular green alga. Polyclonal antibody was raised against a synthetic pentadecameric peptide with an amino acid sequence corresponding to the highly conserved region of the putative frx B proteins of 3 land plants [21]. In this report, protein(s) reacting strongly and specifically with this antibody was detected in the equivalent high-salt extract prepared from purified chloroplast of spinach and tobacco. One strong reaction polypeptide band from tobacco chloroplast was purified from SDS-polyacrylamide gel and subjected to endoproteinase lys C digestion. The resulting polypeptides were separated by reversed-phase chromatography. N-terminal sequencing of 3 purified polypeptides revealed that the protein is encoded by the lsquofrxB genersquo identified from DNA sequence analysis.
Keywords:chloroplast  frx B protein  spinach  tobacco  western blot
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