Isolation and expression of a barley beta-1,3-glucanase isoenzyme II gene.
A beta-1,3-glucanase gene from Hordeum vulgare was isolated by a PCR strategy, cloned and subsequently sequenced. The amplified sequence contained the entire coding region of the isoenzyme II, which is interrupted by a 165 bp intron at 73 bp downstream the starting codon. This intron contains all the elements required for the processing mechanism in monocots: a high A + U content, the appropriate splice sites in the 5' and 3' ends and four typical YUNAN consensus sequences. Transient transformation of wheat protoplasts with the complete beta-1,3-glucanase gene under the control of maize polyubiquitin promoter revealed that the intron sequence was spliced out. The gene was also expressed at high levels, probably due to an enhancer-like sequence found near the 3' end of the intron.
Zanor
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Centro de Estudíos Fotosintéticos y Bioquímicos, Instituto de Biología Molecular y Celular de Rosario, CONICET, Facultad de Ciencias Bioquímicas y Farmacéuticas, UNR, Argentina.
Valle
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Vallejos
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D001483:Base Sequence; D015152:Blotting, Northern; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005091:Exons; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D043326:Glucan 1,3-beta-Glucosidase; D001467:Hordeum / Q000201:enzymology / Q000235:genetics*; D007438:Introns; D007527:Isoenzymes / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011523:Protoplasts / Q000201:enzymology; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014908:Triticum; D014452:Ubiquitins / Q000235:genetics; D003313:Zea mays; D001617:beta-Glucosidase / Q000235:genetics*
D018744:DNA, Plant; D007527:Isoenzymes; D018749:RNA, Plant; D011993:Recombinant Fusion Proteins; D014452:Ubiquitins; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase