ISOLATION OF CDNAS FROM BRASSICA-NAPUS ENCODING THE BIOTIN-BINDING AND TRANSCARBOXYLASE DOMAINS OF ACETYL-COA CARBOXYLASE - ASSIGNMENT OF THE DOMAIN-STRUCTURE IN A FULL-LENGTH ARABIDOPSIS-THALIANA GENOMIC CLONE

被引:15
作者
ELBOROUGH, KM
SWINHOE, R
WINZ, R
KROON, JTM
FARNSWORTH, L
FAWCETT, T
MARTINEZRIVAS, JM
SLABAS, AR
机构
[1] Lipid Molecular Biology Group, Biological Sciences Department, University of Durham, Durham DH1 3LE, South Road
关键词
D O I
10.1042/bj3010599
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
One independent and two overlapping rape cDNA clones have been isolated from a rape embryo library. We have shown that they encode a 2.3 kb and a 2.5 kb stretch of the full-length acetyl-CoA carboxylase (ACCase) cDNA, corresponding to the biotin-binding and transcarboxylase domains respectively. Using the cDNA in Northern-blot analysis we have shown that the mRNA for ACCase has a higher level of expression in rape seed than in rape leaf and has a full length of 7.5 kb. The level of expression during rape embryogenesis was compared with both oil deposition and expression of two fatty acid synthetase components enoyl-(acyl-carrier-protein) reductase and 3-oxoacyl-(acyl-carrier-protein) reductase. Levels of ACCase mRNA were shown to peak at 29 days after anthesis during embryonic development, similarly to enoyl-(acyl-carrier-protein) reductase and 3-oxoacyl-(acyl-carrier-protein) reductase mRNA. In addition, a full-length genomic clone (19 kb) of Arabidopsis ACCase has been isolated and partially sequenced. Analysis of the clone has allowed the first plant ACCase activity domains (biotin carboxylase-biotin binding-transcarboxylase) to be ordered and assigned. Southern-blot analysis using the Arabidopsis clone indicates that ACCase is a single-copy gene in Arabidopsis but is encoded by a small gene family in rape.
引用
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页码:599 / 605
页数:7
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