LOCUS GO546549 631 bp mRNA linear EST 14-APR-2009 DEFINITION Mdfb8004A12_e3507.b1 Apple_EST_Mdfb Malus domestica cDNA 3' similar to gb|AAF97824.1|AC034107_7 Strong similarity to EF-1-alpha-related GTP-binding protein (SUP1) from Nicotiana tabacum gb|L38828 and is a member of the elongation factor Tu PF|00009 family. ESTs gb|W43190, gb|W43332, gb|AI995372, gb|AV563399, gb|AV549134, mRNA sequence. ACCESSION GO546549 VERSION GO546549.1 DBLINK BioSample: SAMN00176549 KEYWORDS EST. SOURCE Malus domestica (apple) ORGANISM Malus domestica Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae; rosids; fabids; Rosales; Rosaceae; Amygdaloideae; Maleae; Malus. REFERENCE 1 (bases 1 to 631) AUTHORS Korban,S., Vodkin,L., Liu,L., Gasic,K., Gonzales,O., Hernandez,A., Aldwinckle,H., Malnoy,M., Carroll,N., Goldsbrough,P., Orvis,K., Clifton,S., Pape,D., Marra,M., Hillier,L., Martin,J., Wylie,T., Dante,M., Theising,B., Bowers,Y., Gibbons,M., Ritter,E., Ronko,I., Tsagareishvili,R., Kennedy,S., Waterston,R. and Wilson,R. TITLE Apple Functional Genomics grant - NSF 0321702 JOURNAL Unpublished COMMENT Contact: Schuyler S. Korban Apple Functional Genomics grant - NSF 0321702 Washington University School of Medicine 4444 Forest Park Parkway, Box 8501, St. Louis, MO 63108, USA Tel: 314 286 1800 Fax: 314 286 1810 Email: submissions@watson.wustl.edu Library material provided by S. Korban/H. Aldwinckle/ M. Malnoy Library constructed by D.O.Gonzalez/L.Vodkin Library sequenced by Washington University Genome Sequencing Center original QR value of 447 was extended to value 447 (,) This trace has been recalled with phred original value before phred recall for SR was 447 Seq primer: -40UP. FEATURES Location/Qualifiers source 1..631 /organism="Malus domestica" /mol_type="mRNA" /cultivar="red delicious" /db_xref="taxon:3750" /tissue_type="Leaves challenged with fire blight (Erwinia amylovora)" /clone_lib="SAMN00176549 Apple_EST_Mdfb" /lab_host="DH10B ampicillin resistant" /note="Vector: pBluescript II SK (+); Site_1: XhoI; Site_2: EcoRI; Total RNA was extracted from freeze dried leaf tissue, using the method described by Wang and Vodkin (Plant Molecular Biology Reporter 12:132-145, 1994). Poly(A)+ mRNA was isolated from total RNA using the PolyATtract mRNA Isolation system III (Promega). The library was prepared using the Stratagene pBluescript II XR cDNA library construction kit. Complementary DNA was synthesized from mRNA using a Poly (dT) sequence with a XhoI restriction site. EcoRI adapters were ligated to the blunt ended cDNA fragments followed by restriction with XhoI. The cDNA insert is protected from XhoI digestion via methylation during first strand cDNA synthesis. The cDNA fragments were directionally cloned into the EcoRI-XhoI restriction site of the pBluescript vector. The ligated cDNA fragments were transformed into E. coli ElectroMax DH10B host cells. Transformation efficiency: 1.30E+05 Average insert Size by PCR: 1420 bp" BASE COUNT 172 a 102 c 124 g 233 t ORIGIN 1 tctttcaagg tcaaggtatt gtttgaaaat ggattttcgg tcattggccc ctatggaaat 61 tatacggagg gggaatgatg agggggggat cattggctcc ctaaaaattt attaaactgt 121 caaagtaacc ccaatttggt gatttattgt taaatgtatt ctctttaatt ttctttaatc 181 ccctgaattc ctttctgtag aatgcctatc ataaacaaat tcaaggactt ggaaacagtt 241 gttatgggca aagtgaaatc tgggtccgtg cgtgaaggga attcattgtt tgtcttgcca 301 aataaggccc cattgaaacc tgttgctata tactttgata aaaatagggg gaaccgtgcg 361 gccccaggtg aaaatctacg ggttaaatta tcaggcattg ataataagaa catattgccg 421 ggctttgttg tgtcaagtgt tggtaagttc tatttcacgg cgtttcactg aattctttgt 481 ccattcttaa aagttttcaa gtattttatt ccttaaaagt aaaaagttgc attgctgctg 541 catgttcgtt ttttcccctt taaagtttga accatttatc tttatttgtt tacagaacct 601 gtcattattt attttatttc tttatttatc a //