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Biochim Biophys Acta ; 1687(1-3): 152-63, 2005 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-15708363

RESUMO

The membrane-bound acyl-CoA elongase complex is a key enzyme responsible for erucoyl-CoA synthesis. Among the four putative genes encoding the four moieties of this complex in Brassica napus seeds, only one has been characterized, the Bn-fae1 gene, which encodes the 3-ketoacyl-CoA synthase. The genes encoding the other enzymes (3-ketoacyl-CoA reductase, 3-hydroxyacyl-CoA dehydratase and trans-2,3-enoyl-CoA reductase) have not been identified. We cloned two 3-ketoacyl-CoA reductase cDNA isoforms, Bn-kcr1 and Bn-kcr2, from B. napus seeds. Their function was identified by heterologous complementation in yeast by restoring elongase activities. The comparison of Bn-kcr mRNA expression in different B. napus tissues showed that the genes were preferentially expressed in seeds and roots. We also investigated the regulation of gene expression in High Erucic Acid Rapeseed (HEAR) and in Low Erucic Acid Rapeseed (LEAR) cultivars during seed development. The co-expression of Bn-fae1 and Bn-kcr observed in HEAR cultivar during seed development was different in LEAR cultivar, suggesting that expression of both genes was directly or indirectly linked.


Assuntos
Oxirredutases do Álcool/metabolismo , Brassica napus , Ácidos Erúcicos/metabolismo , Regulação Enzimológica da Expressão Gênica , Sementes/enzimologia , Sementes/crescimento & desenvolvimento , 3-Oxoacil-(Proteína Carreadora de Acil) Redutase , Oxirredutases do Álcool/genética , Sequência de Aminoácidos , Brassica napus/anatomia & histologia , Brassica napus/química , Brassica napus/enzimologia , Brassica napus/genética , Clonagem Molecular , Ácidos Graxos/biossíntese , Regulação da Expressão Gênica de Plantas , Isoenzimas/genética , Isoenzimas/metabolismo , Dados de Sequência Molecular , Raízes de Plantas/enzimologia , Alinhamento de Sequência
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