GrainGenes Sequence Report: CD663662

[Submit comment/correction]

Sequence

CD663662

External Databases

TIGR Gene Index

NCBI UniGene

DB Remark

Locus Source: Hordeum vulgare subsp. vulgare

UniGene title 'Transcribed locus, moderately similar to NP_001042192.1 Os01g0178500 [Oryza sativa (japonica cultivar-group)]'

Keyword

EST

Species

Hordeum vulgare subsp. vulgare

Cultivar

Dicktoo

Clone Library

Drought-stressed Dicktoo barley epidermis cDNA library

Tissue

lower leaf epidermis

Developmental Stage

1-2 week seedlings

Data Source

genbank	Release 137, Aug 15 2003
genbank	Updated Nov 2006

Title

UCRHV18_06ba04_b1 Drought-stressed Dicktoo barley epidermis cDNA library Hordeum vulgare subsp. vulgare cDNA clone UCRHV18_06ba04, mRNA sequence.

Strain

lab_host E. coli TJC121

vulgare

Clone

UCRHV18_06ba04

Probe

UMB302

Remark

DB_xref: taxon:112509

Feature: source: mol_type = 'mRNA';

Locus Comment: Contact: Timothy J. Close; Department of Botany & Plant Sciences; University of California; Riverside, CA 92521-0124, USA; Tel: 909-787-3318; Fax: 909-787-4437; Email: timothy.close@ucr.edu; Seq primer: T7.

Note: Vector: Lambda Uni-ZAP XR, excised phagemid; Site_1: EcoRI; Site_2: XhoI; Seeds of barley (Hordeum vulgare L. cv. Dicktoo) were germinated in dishpans containing UCR-mix soil. Seedlings were kept in a growth chamber at 20C (day/night) and allowed to grow at 60-70% soil moisture content. After 3-4 days, the water was withheld in order to apply drought until the soil moisture content was reduced to 10-12%, which took another 3-4 days. At the time of extraction of epidermis, leaf water and osmotic potentials and extracted lower epidermis osmotic potentials had dropped to 15.9%, 20.2% and 24.7% of controls, respectively, as determined using a vapor pressure osmometer (Model 5100C, Wescor, Inc., Logan, UT). Epidermal strips were quickly peeled off of seedlings and immediately submerged in liquid nitrogen. About 15-20 g of epidermal tissue was collected and used to extract total RNA. Total RNA was extracted using Concert Plant RNA Reagent (Invitrogen 12322-012). Poly(A) RNA was purified using PolyATrack mRNA Isolation System IV (Promega). A cDNA library was made using a Uni-ZAP cDNA synthesis kit (Stratagene). A total of 1 million primary lambda cDNA clones were mass-excised in vivo to give a population of pBluescript SK(-) phagemids. All steps to this point were performed in the TJ Close lab at the University of California, Riverside by A. Wahid with some assistance from R.D. Fenton. Phagemids were plated on the TJC121 host strain, plasmid DNA purified, cDNA clones archived, and DNA sequences determined using the T7 primer (mainly 3' end reads) using an ABI3730 at the Arizona Genomics Institute, University of Arizona (Collura, Feuerbacher, Kim, Kudrna, Wing, Yu). Chromatogram files were transmitted to UC Riverside (by Yu), then processed at UC Riverside (by Wanamaker) using the HarvEST pipeline (http:

DNA

tttttttttttttttttgcaccatancacaacatgaacttcacccttaac
catcacttggcacacacaaaaaaactcagatttaccaaggcaaaggcaag
caaccacaggtaacctcgcaacgagtgcgcgcagacagcagagagcaccg
agctctgcaagtttggtcttattcatcaacagacgaacaaacaaacagac
agactatatatatatatatatatatatatatatatatatgtagaggtgta
tatttcaagctcccagcactttcaccatcaccatctttctatacgtagta
gtagtagtagtagagagtgctacagacgagtaatgaaaccaacatctttt
cacgactcaagagcatcgggcgaccgaaagcggtaagcgaagataatagc
tgtacacgttcctatctcctcatcagagcaaagccccctcctcctttcag
gcagcaccattcgtaagggccacgcagtgcacctcaccgggcagctccag
gtgcttctctctc


GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture.