starch-branching enzyme gene in wheat produces alternatively spliced transcripts

• Published in: Plant molecular biology Vol. 40; no. 6; pp. 1019 - 1030
• Main Authors: Baga, M, Glaze, S, Mallard, C.S, Chibbar, R.N
• Format: Journal Article
• Language: English
• Published: Netherlands Springer Nature B.V 01-08-1999
• Subjects: 1,4-alpha-Glucan Branching Enzyme - genetics; Alternative Splicing; Amino Acid Sequence; amino acid sequences; Base Sequence; beta-glucuronidase; Deoxyribonucleic acid; DNA; DNA, Plant - chemistry; DNA, Plant - genetics; DNA, Plant - isolation & purification; endosperm; Exons; genbank/aj237897; Gene Expression; Genes, Plant - genetics; genetic transformation; Genetic Variation; hexosyltransferases; histochemistry; Introns; kinases; messenger RNA; Molecular Sequence Data; nucleotide sequences; promoter regions; Proteins; recombinant DNA; Recombinant Fusion Proteins - genetics; RNA, Messenger - genetics; RNA, Plant - genetics; RNA, Plant - metabolism; sbe1 gene; seeds; Seeds - genetics; Sequence Analysis, DNA; signal peptide; Starch; structural genes; Tissue Distribution; Transcription, Genetic; Transformation, Genetic; transgenic plants; Triticum - cytology; Triticum - enzymology; Triticum - genetics; Triticum aestivum; Wheat; zymogens
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1023/A:1006286807176

A wheat gene, denoted Sbe1, encoding a type I starch-branching enzyme (SBEI) was isolated from a genomic library and shown to comprise 14 exons distributed over a 5.7 kb DNA region. Analyses of kernel RNA by 5' rapid amplification of cDNA ends (5'-RACE) and reverse transcription-polymerase chain reaction (RT-PCR) demonstrated a considerable sequence variation at the 5' ends of SBEI gene transcripts. DNA sequence alignments between the 5'-RACE products and the Sbe1 genomic DNA indicated that the first two exons and first intron were differentially processed to generate three classes of the mature transcript. One form of the SBEI gene transcript in 12-day old kernels contained the exon I + II + III combination at the 5' end, whereas other forms differed by inclusion of intron 1 or exclusion of exon II sequences. RT-PCR analysis of Sbe1-uidA::nptII chimeric mRNA produced in transgenic wheat cultured cells confirmed that the isolated Sbe1 was able to produce all three forms of SBEI gene transcripts by alternative splicing of the primary mRNA. The variants of processed Sbe1 mRNA were potentially translated into N-terminal variants of the SBEI precursor with different transit peptide sequences.