Molecular cloning and characterization of the Amylose-Extender gene encoding starch branching enzyme IIB in maize

The amylose-extender (Ae) gene encoding starch-branching enzyme IIb (SBEIIb) in maize is predominantly expressed in endosperm and embryos during kernel development. A maize genomic DNA fragment (-2964 to +20,485) containing the Ae gene was isolated and sequenced. The maize Ae mRNA is derived from 22...

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Bibliographic Details
Published in:Plant molecular biology 1998-12, Vol.38 (6), p.945-956
Main Authors: Kim, K N, Fisher, D K, Gao, M, Guiltinan, M J
Format: Article
Language:English
Subjects:
1,4-alpha-Glucan Branching Enzyme - biosynthesis
1,4-alpha-Glucan Branching Enzyme - genetics
Bacteria
Base Sequence
Cloning
Cloning, Molecular
Conserved Sequence
Corn
Exons
Gene Expression Regulation, Developmental
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Plant
Genes
Genes, Plant
Introns
Luciferases - biosynthesis
Luciferases - genetics
Molecular Sequence Data
Recombinant Fusion Proteins - biosynthesis
Restriction Mapping
Seeds - enzymology
Starch
Zea mays
Zea mays - enzymology
Zea mays - genetics
Zea mays - growth & development
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Summary:The amylose-extender (Ae) gene encoding starch-branching enzyme IIb (SBEIIb) in maize is predominantly expressed in endosperm and embryos during kernel development. A maize genomic DNA fragment (-2964 to +20,485) containing the Ae gene was isolated and sequenced. The maize Ae mRNA is derived from 22 exons distributed over 16,914 bp. Twenty-one introns, differing in length from 76 bp to 4020 bp, all have conserved junction sequences (GT..AG). Sequence analysis of the 5'- and 3'-flanking regions revealed a consensus TATA-box sequence located 28 bp upstream of the transcription initiation site as determined by primer extension analysis, and a putative polyadenylation signal observed 29 bp upstream of the polyadenylation site based on cDNA sequence. Genomic Southern blot analysis suggests that a single Ae gene is present in the maize genome. Promoter activity was confirmed by testing a transcriptional fusion of the Ae 5'-flanking region between -2964 and +100 to a luciferase reporter gene in a transient expression assay using maize endosperm suspension cultured cells. 5' deletion analysis revealed that the 111 bp region from -160 to -50 is essential for high-level promoter activity.
ISSN:0167-4412
1573-5028
DOI:10.1023/A:1006057609995