RESUMO
The aim of this study is to explore the effects of heat stresses on global gene expression profiles and to identify the candidate genes for the heat stress response in commercial baker's yeast (Saccharomyces cerevisiae) by using microarray technology and comparative statistical data analyses. The data from all hybridizations and array normalization were analyzed using the GeneSpringGX 12.1 (Agilent) and the R 2.15.2 program language. In the analysis, all required statistical methods were performed comparatively. For the normalization step, among alternatives, the RMA (Robust Microarray Analysis) results were used. To determine differentially expressed genes under heat stress treatments, the fold-change and the hypothesis testing approaches were executed under various cut-off values via different multiple testing procedures then the up/down regulated probes were functionally categorized via the PAMSAM clustering. The results of the analysis concluded that the transcriptome changes under the heat shock. Moreover, the temperature-shift stress treatments show that the number of differentially up-regulated genes among the heat shock proteins and transcription factors changed significantly. Finally, the change in temperature is one of the important environmental conditions affecting propagation and industrial application of baker's yeast. This study statistically analyzes this affect via one-channel microarray data.
Assuntos
Regulação Fúngica da Expressão Gênica , Resposta ao Choque Térmico , Saccharomyces cerevisiae/genética , Transcriptoma , Biologia Computacional , Perfilação da Expressão Gênica , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/fisiologia , Análise de Sequência de RNARESUMO
Two problems that dog current microarrays analyses are (i) the relatively arbitrary nature of data preprocessing and (ii) the inability to incorporate spot quality information in inference except by all-or-nothing spot filtering. In this chapter we propose an approach based on using weights to overcome these two problems. The first approach uses weighted p-values to make inference robust to normalization and the second approach uses weighted spot intensity values to improve inference without any filtering.
Assuntos
Perfilação da Expressão Gênica/métodos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Algoritmos , Animais , Interpretação Estatística de Dados , Humanos , Camundongos , Modelos Estatísticos , Reconhecimento Automatizado de Padrão , RNA Mensageiro/metabolismo , SoftwareRESUMO
We consider a new frequentist gene expression index for Affymetrix oligonucleotide DNA arrays, using a similar probe intensity model as suggested by Hein and others (2005), called the Bayesian gene expression index (BGX). According to this model, the perfect match and mismatch values are assumed to be correlated as a result of sharing a common gene expression signal. Rather than a Bayesian approach, we develop a maximum likelihood algorithm for estimating the underlying common signal. In this way, estimation is explicit and much faster than the BGX implementation. The observed Fisher information matrix, rather than a posterior credibility interval, gives an idea of the accuracy of the estimators. We evaluate our method using benchmark spike-in data sets from Affymetrix and GeneLogic by analyzing the relationship between estimated signal and concentration, i.e. true signal, and compare our results with other commonly used methods.