Isolation and characterization of cDNA encoding P-19.5 protein accumulated preferentially at early stage of carrot somatic embryogenesis
We found P-19.5 protein that preferentially accumulated in cell clusters and globular embryos at the early stage of carrot somatic embryogenesis. P-19.5 protein was isolated from 9-day-old cell clusters with SDS-PAGE, and then determined the amino acid sequences of a N-terminal peptide and some inte...
Gespeichert in:
Veröffentlicht in: | Plant science (Limerick) 2004-12, Vol.167 (6), p.1211-1217 |
---|---|
Hauptverfasser: | , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
Zusammenfassung: | We found P-19.5 protein that preferentially accumulated in cell clusters and globular embryos at the early stage of carrot somatic embryogenesis. P-19.5 protein was isolated from 9-day-old cell clusters with SDS-PAGE, and then determined the amino acid sequences of a N-terminal peptide and some internal peptides. Using each primer for these peptides, the full-length sequence of cDNA encoding P-19.5 protein was determined by analyzing 3′-RACE and 5′-RACE products. We regard P-19.5 protein as an allergen-like protein because the P-19.5 protein sequence predicted from cDNA was homologous to celery major allergen (75%), carrot major allergen (70–72%),
Pimpinella brachycarpa pathogenesis-related protein (71%), and parsley pathogenesis-related protein (63%). We found concomitantly two separate cDNAs (
P-16 and
P-19.5 H cDNAs) during analyzing
P-19.5 cDNA:
P-16 cDNA was cloned and isolated from P-16 protein that was recognized against P-19.5 protein antiserum, and
P-19.5 H cDNA that had homology (80.5% amino acid identity) with
P-19.5 cDNA was detected during analyzing
P-19.5 cDNA. Northern blotting revealed that
P-19.5 and
P-19.5 H transcripts were highly expressed at early stages of cell cluster and globular embryos, whereas
P-16 transcript was invariably expressed at all stages from cell clusters to plantlets. |
---|---|
ISSN: | 0168-9452 1873-2259 |
DOI: | 10.1016/j.plantsci.2004.05.025 |