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.
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.
Elborough
K M
KM
Biological Sciences Department, University of Durham, U.K.
Swinhoe
R
R
Winz
R
R
Kroon
J T
JT
Farnsworth
L
L
Fawcett
T
T
Martinez-Rivas
J M
JM
Slabas
A R
AR
D000103:Acetyl-CoA Carboxylase / Q000737:chemistry* / Q000235:genetics*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D001665:Binding Sites; D001710:Biotin / Q000378:metabolism*; D015152:Blotting, Northern; D015139:Blotting, Southern; D001937:Brassica / Q000196:embryology / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification*; D005227:Fatty Acids / Q000032:analysis; D015870:Gene Expression; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D012639:Seeds / Q000737:chemistry / Q000235:genetics
D018076:DNA, Complementary; D005227:Fatty Acids; D012333:RNA, Messenger; D001710:Biotin; D000103:Acetyl-CoA Carboxylase