Intron-mediated enhancement of heterologous gene expression in maize

Chimeric genes containing the coding sequence for bacterial chloramphenicol acetyl transferase (CAT) have been introduced by electroporation into maize protoplasts (Black Mexican Sweet) and transient expression monitored by enzyme assays. Levels of CAT expression were enhanced 12-fold and 20-fold re...

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Bibliographic Details
Published in:Plant molecular biology 1990-12, Vol.15 (6), p.913-920
Main Authors: Mascarenhas, D. (Sandoz Crop Protection, Palo Alto, CA (USA)), Mettler, I.J, Pierce, D.A, Lowe, H.W
Format: Article
Language:English
Subjects:
Alcohol Dehydrogenase - genetics
Bacterial Proteins - genetics
Base Sequence
Biological and medical sciences
Chloramphenicol O-Acetyltransferase - biosynthesis
Chloramphenicol O-Acetyltransferase - genetics
DNA - genetics
EXPRESION GENICA
EXPRESSION DES GENES
Fundamental and applied biological sciences. Psychology
GENE EXPRESSION
Gene Expression Regulation
Genes, Bacterial
Genes, Plant
Genes, Synthetic
Genetic Vectors
Introns
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Plant Proteins - genetics
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - genetics
Regulatory Sequences, Nucleic Acid
Transformation, Genetic
ZEA MAYS
Zea mays - genetics
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Summary:Chimeric genes containing the coding sequence for bacterial chloramphenicol acetyl transferase (CAT) have been introduced by electroporation into maize protoplasts (Black Mexican Sweet) and transient expression monitored by enzyme assays. Levels of CAT expression were enhanced 12-fold and 20-fold respectively by the inclusion of maize alcohol dehydrogenase-1 introns 2 and 6 in the chimeric construct. This enhancement was seen when the intron was placed within the 5' translated region but not when it was located upstream of the promoter or within the 3' untranslated region. Deletion of exon sequences adjacent to intron 2 abolished its ability to mediate enhancement of CAT gene expression. Northern analysis of protoplasts electroporated with intron constructs revealed elevated levels of CAT mRNA. However, this elevation was insufficient to account for the increased enzyme activity. One explanation of these results is that splicing affects both the quantity and quality of mRNA.
ISSN:0167-4412
1573-5028
DOI:10.1007/BF00039430