Molecular cloning and characterization of OsPSK, a gene encoding a precursor for phytosulfokine-α, required for rice cell proliferation

Molecular cloning and characterization of OsPSK, a gene encoding a precursor for... We previously characterized an OsPSK cDNA encoding a precursor of phytosulfokine-α (PSK-α), a peptide plant growth factor. Southern blot analysis suggested that OsPSK is a single-copy gene in rice, which we have isolated and characterized. The OsPSK gene consists of one large intron and two exons. The 5-amino acid PSK-α sequence located close to the COOH-terminus of the precursor is encoded in the second exon. A putative TATA box was found at position −68 with respect to the transcription initiation site. Upstream of this sequence, several potential regulatory elements, including one CAAT-box, three CCAAT-boxes, one enhancer core-like sequence, and three E-boxes could be identified. By constructing plasmids with various lengths of the 5′-upstream regions of the OsPSK gene fused to the coding sequence for bacterial β-glucuronidase (GUS), we demonstrated a region 1.9 kb upstream of the transcription initiation point, which contains most of the putative 5′-regulatory elements, to be sufficient for maximal-level GUS expression in transformed rice Oc cells. The promoter of the OsPSK gene gave significantly higher levels of GUS expression than the CaMV 35S promoter. These results suggest that the OsPSK promoter could be useful for the constitutive expression of a foreign gene at high levels in transformed rice culture cells. Northern blot analyses suggest that the expression of OsPSK is reinforced by auxin and cytokinin. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Molecular cloning and characterization of OsPSK, a gene encoding a precursor for phytosulfokine-α, required for rice cell proliferation

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Publisher
Kluwer Academic Publishers
Copyright
Copyright © 2000 by Kluwer Academic Publishers
Subject
Life Sciences; Biochemistry, general; Plant Sciences; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1023/A:1026576423870
Publisher site
See Article on Publisher Site

Abstract

We previously characterized an OsPSK cDNA encoding a precursor of phytosulfokine-α (PSK-α), a peptide plant growth factor. Southern blot analysis suggested that OsPSK is a single-copy gene in rice, which we have isolated and characterized. The OsPSK gene consists of one large intron and two exons. The 5-amino acid PSK-α sequence located close to the COOH-terminus of the precursor is encoded in the second exon. A putative TATA box was found at position −68 with respect to the transcription initiation site. Upstream of this sequence, several potential regulatory elements, including one CAAT-box, three CCAAT-boxes, one enhancer core-like sequence, and three E-boxes could be identified. By constructing plasmids with various lengths of the 5′-upstream regions of the OsPSK gene fused to the coding sequence for bacterial β-glucuronidase (GUS), we demonstrated a region 1.9 kb upstream of the transcription initiation point, which contains most of the putative 5′-regulatory elements, to be sufficient for maximal-level GUS expression in transformed rice Oc cells. The promoter of the OsPSK gene gave significantly higher levels of GUS expression than the CaMV 35S promoter. These results suggest that the OsPSK promoter could be useful for the constitutive expression of a foreign gene at high levels in transformed rice culture cells. Northern blot analyses suggest that the expression of OsPSK is reinforced by auxin and cytokinin.

Journal

Plant Molecular BiologySpringer Journals

Published: Oct 16, 2004

References

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