cDNA cloning, mRNA expression, and mutational analysis of the squalene synthase gene of Lotus japonicus

被引:27
作者
Akamine, S
Nakamori, K
Chechetka, SA
Banba, M
Umehara, Y
Kouchi, H
Izui, K
Hata, S [1 ]
机构
[1] Kyoto Univ, Grad Sch Biostudies, Lab Plant Physiol, Sakyo Ku, Kyoto 6068502, Japan
[2] Kyoto Univ, Grad Sch Agr, Div Appl Biosci, Sakyo Ku, Kyoto 6068502, Japan
[3] Natl Inst Agrobiol Sci, Tsukuba, Ibaraki 3058602, Japan
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION | 2003年 / 1626卷 / 1-3期
关键词
in situ hybridization; recombinant protein; site-directed mutagenesis; Lotus japonicus;
D O I
10.1016/S0167-4781(03)00042-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A full-length cDNA for squalene synthase was isolated from Lotus japonicus, a model leguminous plant. The transcript was abundant in roots, symbiotic root nodules, and shoots, in that order. In situ hybridization revealed that the mRNA level is high in expanding root cells but low in dividing root tip ones. The transcript is also abundant in vascular bundles and the basal portions of mature nodules. L. japonicus squalene synthase has an unusual Asp residue near the active site, where mammalian enzymes have Gin, and replacement of the Gln by Glu has been reported to cause severe inactivation. Site-directed mutagenesis of the L. japonicus enzyme and assaying in vitro showed that this Asp residue can be substituted by not only Gln but also Glu, suggesting that the local structure of plant squalene synthases is different from that of mammalian enzymes. (C) 2003 Elsevier Science B.V All rights reserved.
引用
收藏
页码:97 / 101
页数:5
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