The combination of a novel stimulatory element in the first exon of the maize Shrunken-1 gene with the following intron 1 enhances reporter gene expression up to 1000-fold.
Both exon 1 and intron 1 of the maize Shrunken-1 (Sh1) gene individually stimulate expression of reporter genes in transient gene expression experiments if present within the transcription unit. The Sh1 exon 1 mediates a 10-fold increase in activity when inserted at the 5' end of the bacterial chloramphenicol transacetylase (CAT) marker gene in both monocot and dicot protoplasts. The Sh1 intron 1 enhances chimeric gene expression in rice and maize protoplasts approximately 100-fold but inhibits CAT expression in tobacco protoplasts. In combination, the stimulatory effects of Sh1 exon 1 and intron 1 are multiplicative in monocot protoplasts resulting in a final enhancement of up to 1000-fold compared to the unmodified CAT or luciferase marker genes.
Maas
C
C
Institut für Genetik Universität zu Köln, Germany.
Laufs
J
J
Grant
S
S
Korfhage
C
C
Werr
W
W
D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA; D005091:Exons; D005819:Genetic Markers / Q000235:genetics; D007438:Introns; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011523:Protoplasts; D012045:Regulatory Sequences, Nucleic Acid; D014026:Nicotiana / Q000235:genetics; D003313:Zea mays / Q000235:genetics*
D005819:Genetic Markers; D004247:DNA; D008156:Luciferases; D015500:Chloramphenicol O-Acetyltransferase