期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 118, 期 -, 页码 42-50出版社
ELSEVIER
DOI: 10.1016/j.mimet.2015.08.015
关键词
qRT-PCR; Talaromyces marneffei; Penicillium marneffei; Actin; Reference gene
资金
- National Research University Project under Thailand's Office of the Higher Education Commission
- Faculty of Medicine, Chiang Mai University
Talaromyces marneffei (or Penicillium marneffei) is an opportunistic pathogen that can cause disseminated disease in human immunodeficiency virus (HIV)-infected patients, especially in Southeast Asia. T. marneffei is a thermally dimorphic fungus. Typically, T. marneffei has an adaptive morphology. It grows in a filamentous form (mould) at 25 degrees C and can differentiate to produce asexual spores (conidia). In contrast, at 37 degrees C, it grows as yeast cells that divide by fission. This study aimed to validate a quantitative reverse-transcription polymerase chain reaction (qRT-PCR) for gene expression analysis in T. marneffei. Analysis of relative gene expression by qRT-PCR requires normalization of data using a proper reference gene. However, suitable reference genes have not been identified in gene expression studies across different cell types or under different experimental conditions in T. marneffei. In this study, four housekeeping genes were selected for analysis: beta-actin (act); glyceraldehyde-3-phosphate dehydrogenase (gapdh); beta-tubulin (benA) and 18S rRNA. Two analysis programs; BestKeeper and geNorm software tools were used to validate the expression of the candidate normalized genes. The results indicated that the actin gene was the one which was most stably expressed and was recommended for use as the endogenous control for gene expression analysis of all growth forms in T. marneffei by qRT-PCR under normal and stress conditions. (C) 2015 Elsevier B.V. All rights reserved.
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