Complete structure of the gene for phosphoenolpyruvate carboxylase from maize

• Published in: European journal of biochemistry Vol. 181; no. 3; pp. 593 - 598
• Main Authors: Matsuoka, M. (National Institute of Agrobiological Resources, Tsukuba Science City, Ibaraki), Minami, E.I
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 15.05.1989; Blackwell
• Subjects: Base Sequence; Biological and medical sciences; Blotting, Southern; Carboxy-Lyases - genetics; Cloning, Molecular; Exons; Fundamental and applied biological sciences. Psychology; GENE; GENES; Genes. Genome; GENOMAS; GENOME; GENOMES; Introns; LIASAS; LYASE; LYASES; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Nucleic Acid Hybridization; Peptide Mapping; Phosphoenolpyruvate Carboxylase - genetics; Promoter Regions, Genetic; Transcription, Genetic; ZEA MAYS; Zea mays - enzymology; Monocotyledones; Zea mays; Radiolabelling; Molecular structure; Nucleotide sequence; Gel electrophoresis; Copy number; Gene; Gramineae; Angiospermae; Multigene family; Regulatory sequence; Spermatophyta; Molecular cloning; Southern blotting; Transcription initiation; Phosphoenolpyruvate carboxylase
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1111/j.1432-1033.1989.tb14765.x

Phosphoenolpyruvate carboxylase is a key enzyme in photosynthesis in some plants that exploit the C4 photosynthetic pathway for the fixation of CO2. We cloned the gene for this enzyme from maize genomic libraries and analyzed its complete primary structure. The sequence of the cloned gene spans 6781 bp and consists of 10 exons and 9 introns. The site of initiation of transcription is located 84 nucleotides upstream from the first nucleotide of the initiation codon (position –84), as determined by the method of primer‐extension analysis. The analysis suggests that there is another initiation site located at position –81. The 5′‐flanking region of the gene lacks typical TATA and CCAAT elements in the anticipated regions, but there is a TATA‐similar sequence (TATTT) around the –30 regions as well as sequence homologous to the Sp‐1 protein‐binding site (CCGCCC). Six long, direct repeated sequences and a light‐responsive element are also present in the 5′‐flanking region. The results of Southern blot analysis indicated that the phosphoenolpyruvate carboxylase gene exists as a small multi‐gene family, but the enzyme that is expressed at high levels in green leaves and is involved in C4 photosynthesis is encoded by a single‐copy gene in the maize genome.