Molecular structure and function of the porcine arachidonate 12-lipoxygenase gene.
The gene encoding arachidonate 12-lipoxygenase was cloned from a porcine EMBL3 genomic library using a cDNA probe of the enzyme, and its nucleotide sequence was determined. The gene consists of 14 exons with 13 introns, and spans approximately 8 kilobases. Analysis of splice junctions indicated that all of the splice donor and acceptor sites conformed to the GT/AG rule. An approximately 1-kilobase region upstream of the coding sequence contains nine GC-boxes for potential Sp1-binding sites at positions -77, -135, -145, -165, -214, -636, -643, -684, and -813. There are two sets of AP-2 binding sequence at positions -234 and -402. Neither typical TATA box nor CCAAT box is found in this region. The transcriptional start site was determined by primer extension analysis, and was tentatively identified as a cytidine residue located 19 bases upstream from initiation codon. Southern blot analysis revealed the presence of one copy of 12-lipoxygenase gene per haploid genome. We found striking similarities in genomic organization as well as the promoter sequences between the porcine 12-lipoxygenase and the rabbit 15-lipoxygenase genes, suggesting that these genes are evolutionarily related.
Arakawa
T
T
Department of Biochemistry, Tokushima University School of Medicine, Japan.
Oshima
T
T
Kishimoto
K
K
Yoshimoto
T
T
Yamamoto
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D004247:DNA / Q000235:genetics; D005091:Exons; D007438:Introns; D008969:Molecular Sequence Data; D015394:Molecular Structure; D012326:RNA Splicing; D012689:Sequence Homology, Nucleic Acid; D013552:Swine; D014158:Transcription, Genetic
D004247:DNA; D001092:Arachidonate 12-Lipoxygenase