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

Kyung Nam Kim, Dane K. Fisher, Ming Gao, Mark J. Guiltinan

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

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 +20485) containing the Ae gene was isolated and sequenced. The maize Ae mRNA is derived from 22 exons distributed over 16914 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.

Original languageEnglish (US)
Pages (from-to)945-956
Number of pages12
JournalPlant molecular biology
Volume38
Issue number6
DOIs
StatePublished - Dec 1 1998

Fingerprint

1,4-alpha-glucan branching enzyme
Amylose
Molecular Cloning
amylose
Zea mays
molecular cloning
corn
Endosperm
Polyadenylation
Genes
5' Flanking Region
genes
endosperm
promoter regions
3' Flanking Region
genomics
TATA box
TATA Box
Conserved Sequence
Transcription Initiation Site

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

Cite this

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title = "Molecular cloning and characterization of the Amylose-Extender gene encoding starch branching enzyme IIb in maize",
abstract = "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 +20485) containing the Ae gene was isolated and sequenced. The maize Ae mRNA is derived from 22 exons distributed over 16914 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.",
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Molecular cloning and characterization of the Amylose-Extender gene encoding starch branching enzyme IIb in maize. / Kim, Kyung Nam; Fisher, Dane K.; Gao, Ming; Guiltinan, Mark J.

In: Plant molecular biology, Vol. 38, No. 6, 01.12.1998, p. 945-956.

Research output: Contribution to journalArticle

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