GrainGenes Sequence Report: EB513867

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Sequence

EB513867

External Databases

TIGR Gene Index

NCBI UniGene

DB Remark

Locus Source: Triticum aestivum (bread wheat)

UniGene title 'Transcribed locus, strongly similar to NP_850807.2 DEAD/DEAH box helicase, putative (RH15) [Arabidopsis thaliana]'

Keyword

EST

Species

Triticum aestivum

Cultivar

Chinese Spring addition line containing chromosome 7E from Thinopyrum elongatum

Clone Library

Ta09_AAFC_ECORC_Fusarium_graminearum_inculated _wheat_heads

Tissue

spikelets

Developmental Stage

anthesis

Data Source

genbank

Updated Nov 2006

Title

Ta09_02f19_R Ta09_AAFC_ECORC_Fusarium_graminearum_inculated_wheat_heads Triticum aestivum cDNA clone Ta09_02f19, mRNA sequence.

Strain

lab_host E. coli Sure Cells

Clone

Ta09_02f19

Probe

MAG2048

Remark

DB_xref: taxon:4565

Feature: source: mol_type = 'mRNA';

Locus Comment: Contact: Ouellet, Therese; Eastern Cereal and Oilseed Research Centre; Agriculture and Agri-food Canada; Neatby Bldg., Central Experimental Farm, Ottawa, Ontario, K1A 0C6,; CANADA; Tel: (613) 759-1658; Fax: (613) 759-1701; Email: ouellettr@agr.gc.ca.

Note: Vector: pGem-T easy; Site_1: EcoRI; Site_2: EcoRI; Wheat plants (Chinese Spring and Chinese Spring addition line containing chromosome 7E from Thinopyrum elongatum) were grown under a 16 h light (20oC) /8 h dark (16oC) regime until mid-anthesis. Heads at mid-anthesis of the addition line were point-inoculated with Fg spores (10 ul of 1 X 105 spores/ml) at every fully developed spikelet. Water inoculation was used on Chinese Spring plants. Inoculated spikelets (with Fg or water) were harvested immediately after inoculation. Total RNA was extracted using guanidinium thiocyanate followed by pelleting through CsCl2. Poly(A+)RNA was isolated following the manufacturer's instructions using MPG Streptavidin Biotinylated Oligo (dT)25 Complex (CPG, NJ). A subtraction suppression hybridization (SSH) library was generated with the PCR-Select cDNA Subtraction Kit (BD Biosciences, Missisauga, ON, Canada), using cDNA from F. graminearum-inoculated tissues of the addition line as the tester, subtracted with water-inoculated cDNA from Chinese Spring as the driver. The enriched SSH PCR products were cloned into the pGEM T-easy vector (Promega).

DNA

acagctgctgctcctgctgcctccctcccttggcagcagggcagggcagg
gcaggcaaaaaacagcatcacttcaagcaaaacattcaaggattccatct
tckcsktcsgtcctccttgcaatgtcggcgcgggtctacgacggcatgta
tgttgaagtgtcaatctgctcaggcagctccttgatgtccacctcaaacc
gttcttgcacctggttaaggacgtccgagtccgaggccgacgaaacaaaa
gttatggcgagtcccttggtgccaaaacgcccagcccttccaaccctgtg
caagtaggtatcaggkgaatcaggcatgtcatagtttatcacaatgttga
cacgctcaatgtcgatccctctgccaaccaaatccgtcgccacaagaatc
ctcttgtgcccttccttgaagttcttgtatcgggtcagcctctcttcctg
ggtcatgccagaatggrtgcagattgaagggaagttgcattcacaaagca
acttgttcagctctgcagccctgctaacgctcttcacaaatatcacaact
tggttgaaatcgagagcatccaagagatcgttcaattttcggttcttctc
tgactctgtcaattttatgtagtgctgaacaagaccgtgaagggkcaact
ttgcctcatcatcaacataa


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