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

Biochem J. 1994 Jul 15;301 ( Pt 2)(Pt 2):599-605. doi: 10.1042/bj3010599.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyl-CoA Carboxylase / chemistry*
  • Acetyl-CoA Carboxylase / genetics*
  • Amino Acid Sequence
  • Arabidopsis / genetics*
  • Binding Sites
  • Biotin / metabolism*
  • Blotting, Northern
  • Blotting, Southern
  • Brassica / embryology
  • Brassica / enzymology
  • Brassica / genetics*
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • DNA, Complementary / isolation & purification*
  • Fatty Acids / analysis
  • Gene Expression
  • Molecular Sequence Data
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism
  • Seeds / chemistry
  • Seeds / genetics

Substances

  • DNA, Complementary
  • Fatty Acids
  • RNA, Messenger
  • Biotin
  • Acetyl-CoA Carboxylase

Associated data

  • GENBANK/X77374
  • GENBANK/X77375
  • GENBANK/X77376
  • GENBANK/X77377
  • GENBANK/X77378
  • GENBANK/X77379
  • GENBANK/X77380
  • GENBANK/X77381
  • GENBANK/X77382