Expression of Pisum sativum SAD polypeptides in production hosts and in planta: Tetrameric organization of the protein

被引：2
|
作者：
Scherbak, Nikolai [1 ,2 ]
Brosche, Mikael [3 ]
Ala-Haivala, Anneli [1 ,2 ]
Strid, Hija [2 ,4 ]
Ohrfelt, Annika [3 ,5 ]
Nilsson, Fredrik [5 ]
Strid, Ake [1 ,2 ]
机构：
[1] Univ Orebro, Dept Sci, SE-70182 Orebro, Sweden
[2] Univ Orebro, Orebro Life Sci Center, SE-70182 Orebro, Sweden
[3] Univ Gothenburg, Dept Chem, SE-40530 Gothenburg, Sweden
[4] Univ Orebro, Dept Clin Med, SE-70182 Orebro, Sweden
[5] Dept Bioanalyt Chem, SE-43121 Molndal, Sweden
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2009年 / 63卷 / 01期
关键词：
Environmental stress; Short-chain alcohol dehydrogenase; Short-chain dehydrogenase/reductase; Tetramer; Ultraviolet-B radiation; ULTRAVIOLET-B-RADIATION; MESSENGER-RNA; SECOISOLARICIRESINOL DEHYDROGENASE; FUNCTIONAL ASSIGNMENTS; GENE-EXPRESSION; CHAIN; METABOLISM; CLONING; ENZYME; IDENTIFICATION;
D O I：
10.1016/j.pep.2008.09.004
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
In Pisum sativum, the short-chain alcohol dehydrogenase-like protein (SAD) gene family consists of at least three members (SAD-A, -B, and -C). Expression of two of these genes (SAD-A and -C) in Escherichia coli or Pichia pastoris resulted in full-length soluble proteins. Purified SAD-A was used as antigen for antibody production in rabbits. With these antibodies the recombinant SAD-C protein (which was most highly expressed of the two isoforms) was shown to be a tetramer consisting of a dinner of dimers. The SAD genes are transiently expressed in plants by short exposures to ultraviolet-B radiation (UV-B), as judged by northern blotting. In turn, mRNA accumulation leads to formation of SAD protein in leaf and stem tissue upon prolonged UV-B irradiation. (c) 2008 Elsevier Inc. All rights reserved.
引用
收藏
页码：18 / 25
页数：8
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