Biomedical subjects

Christine D Smart

Publications and source records attributed to Christine D Smart.

4 recordsLinked to original sources

PubMed2005

Bayesian normalization and identification for differential gene expression data.

Commonly accepted intensity-dependent normalization in spotted microarray studies takes account of measurement errors in the differential expression ratio but ignores measurement errors in the total intensity, although the definitions imply the same measurement error components are involved in both statistics. Furthermore, identification of differentially expressed genes is usually considered separately following normalization, which is statistically problematic. By incorporating the measurement errors in both total intensities and differential expression ratios, we propose a measurement-error model for intensity-dependent normalization and identification of differentially expressed genes. This model is also flexible enough to incorporate intra-array and inter-array effects. A Bayesian framework is proposed for the analysis of the proposed measurement-error model to avoid the potential risk of using the common two-step procedure. We also propose a Bayesian identification of differentially expressed genes to control the false discovery rate instead of the ad hoc thresholding of the posterior odds ratio. The simulation study and an application to real microarray data demonstrate promising results.

Bayes Theorem↗

PubMed2004

Core promoter structure in the oomycete Phytophthora infestans.

We have investigated the core promoter structure of the oomycete Phytophthora infestans. The transcriptional start sites (TSS) of three previously characterized P. infestans genes, Piexo1, Piexo3, and Piendo1, were determined by primer extension analyses. The TSS regions were homologous to a previously identified 16-nucleotide (nt) core sequence that overlaps the TSS in most oomycete genes. The core promoter regions of Piexo1 and Piendo1 were investigated by using a transient protoplast expression assay and the reporter gene beta-glucuronidase. Mutational analyses of the promoters of Piexo1 and Piendo1 showed that there is a putative core promoter element encompassing the TSS (-2 to +5) that has high sequence and functional homology to a known core promoter element present in other eukaryotes, the initiator element (Inr). Downstream and flanking the Inr is a highly conserved oomycete promoter region (+7 to +15), hereafter referred to as FPR (flanking promoter region), which is also important for promoter function. The importance of the 19-nt core promoter region (Inr and FPR) in Piexo1 and Piendo1 was further investigated through electrophoretic mobility shift assays (EMSA). The EMSA studies showed that (i) both core promoters were able to specifically bind a protein or protein complex in a P. infestans whole-cell protein extract and (ii) the same mutations that reduced binding of the EMSA complex also reduced beta-glucuronidase (GUS) levels in transient expression assays. The consistency of results obtained using two different assays (GUS transient assays [in vivo] and EMSA studies [in vitro]) supports a convergence of inference about the relative importance of specific nucleotides within the 19-nt core promoter region.

Base Sequence↗

PubMed2003

Characterization of 1,3-beta-glucanase and 1,3;1,4-beta-glucanase genes from Phytophthora infestans.

Three putative exo-1,3-beta-glucanase genes (Piexo1, Piexo2, Piexo3), one endo-1,3-beta-glucanase (Piendo1) and one endo-1,3;1,4-beta-glucanase (Piendo2) gene were cloned and characterized from the oomycete Phytophthora infestans. Southern hybridization revealed that Piexo1, Piexo2, Piexo3, and Piendo2 are single copy, and that Piendo1 is encoded by two copies. Furthermore, the analyses showed that for each gene, one or two closely related gene family members were present. The genes contain no introns. Nucleotide sequence analysis of the promoter regions (200nt upstream of ATG start codon) showed that the regions have 56-81% similarity to a 16-nt core sequence hypothesized to be the initiation of transcription point in oomycetes. The predicted molecular weights (32-83kDa), iso-electric points (4.2-6.7) and amino acid sequences of the five proteins are diverse. All the genes are expressed in in vitro grown mycelia and sporangia, as well as during infection of potatoes. Further, Piendo1 and Piendo2 are also expressed in germinating cysts, and Piendo2 in zoospores.

Gene Dosage↗

PubMed2003

Comparative analyses of potato expressed sequence tag libraries.

The cultivated potato (Solanum tuberosum) shares similar biology with other members of the Solanaceae, yet has features unique within the family, such as modified stems (stolons) that develop into edible tubers. To better understand potato biology, we have undertaken a survey of the potato transcriptome using expressed sequence tags (ESTs) from diverse tissues. A total of 61,940 ESTs were generated from aerial tissues, below-ground tissues, and tissues challenged with the late-blight pathogen (Phytophthora infestans). Clustering and assembly of these ESTs resulted in a total of 19,892 unique sequences with 8,741 tentative consensus sequences and 11,151 singleton ESTs. We were able to identify a putative function for 43.7% of these sequences. A number of sequences (48) were expressed throughout the libraries sampled, representing constitutively expressed sequences. Other sequences (13,068, 21%) were uniquely expressed and were detected only in a single library. Using hierarchal and k means clustering of the EST sequences, we were able to correlate changes in gene expression with major physiological events in potato biology. Using pair-wise comparisons of tuber-related tissues, we were able to associate genes with tuber initiation, dormancy, and sprouting. We also were able to identify a number of characterized as well as novel sequences that were unique to the incompatible interaction of late-blight pathogen, thereby providing a foundation for further understanding the mechanism of resistance.

Cluster Analysis↗