DEVELOPING EXPRESSED SEQUENCED TAGS (ESTS) FOR WATERMELON FRUIT

A cDNA library was assembled using mRNA of watermelon fruit. The cDNA library was normalized and subtracted by hybridization with leaf cDNA of the same watermelon cultivar (Illini Red). 1,046 cDNA clones were sequenced to identify genes associated with fruit development and quality. Of 1,046 cDNA clones sequenced, 832 were unique sequences and designated as expressed sequenced tags (ESTs). Of the 832 ESTs, 205 (24.6%) have not been reported in any other plant species. Additionally, 186 ESTs (22.4%) correspond to genes with unknown function, while 441 ESTs (53.0%) correspond to genes with known function in other plant species. These ESTs are mainly associated with primary metabolism, membrane transport, cytoskeleton synthesis and structure, cell wall and cell division, signal transduction, nucleic acid binding and transcription factors, and defense and stress response. Differential expression of the ESTs was examined using microarray analysis. About 200 (24%) of the 832 ESTs showed differential expression during the development and ripening of watermelon fruit. The ESTs were also screened for simple sequence repeat (SSR) motifs. Of 832 ESTs screened, 177 contain SSR motifs. Primer pairs are being designed for these ESTs, and will be used for development of EST-SSR markers and for mapping on a genetic linkage map constructed for watermelon. This study provides valuable information on genes controlling watermelon fruit development and quality.

Overexpression of the ETS-Related Gene, ERG, Predicts a Worse Outcome in Acute Myeloid Leukemia With Normal Karyotype: A Cancer and Leukemia Group B Study

Purpose To test the prognostic significance of ETS-related gene (ERG) expression in cytogenetically normal primary acute myeloid leukemia (AML). Patients and Methods Pretreatment blood samples from 84 cytogenetically normal AML patients aged less than 60 years, who were characterized for BAALC expression, FLT3 internal tandem duplication (ITD), and MLL partial tandem duplication (PTD) and uniformly treated on Cancer and Leukemia Group B 9621 protocol, were analyzed for ERG expression by real-time reverse transcriptase polymerase chain reaction. Patients were divided into quartiles according to ERG levels and were compared for clinical outcome. High-density oligonucleotide arrays were used to identify genes differentially expressed between high and low ERG expressers. Results With a median follow-up of 5.7 years, patients with the upper 25% of ERG expression values had a worse cumulative incidence of relapse (CIR; P < .001) and overall survival (OS; P = .011) than the remaining patients. In a multivariable analysis, high ERG expression (P < .001) and the presence of MLL PTD (P = .027) predicted worse CIR. With regard to OS, an interaction was observed between expression of ERG and BAALC (P = .013), with ERG overexpression predicting shorter survival only in low BAALC expressers (P = .002). ERG overexpression was an independent prognostic factor even when the unfavorable group of FLT3 ITD patients lacking an FLT3 wild-type allele was included. High ERG expression was associated with upregulation of 112 expressed-sequenced tags and named genes, many of which are involved in cell proliferation, differentiation, and apoptosis. Conclusion ERG overexpression in AML patients with normal cytogenetics predicts an adverse clinical outcome and seems to be associated with a specific molecular signature.