Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis
Braz. j. microbiol
; 47(1): 259-265, Jan.-Mar. 2016. tab, graf
Article
in En
| LILACS
| ID: lil-775129
Responsible library:
BR1.1
ABSTRACT
Abstract The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.
Key words
Full text:
1
Collection:
01-internacional
Database:
LILACS
Main subject:
Reference Standards
/
Entomophthorales
/
Gene Expression Profiling
/
Real-Time Polymerase Chain Reaction
/
Genes, Fungal
Type of study:
Prognostic_studies
Language:
En
Journal:
Braz. j. microbiol
Journal subject:
MICROBIOLOGIA
Year:
2016
Document type:
Article
/
Project document
Affiliation country: