Validation of reference genes for reverse transcription real-time quantitative PCR analysis in the deep-sea bacterium Shewanella psychrophila WP2 | Zendy

Shunzhang Liu | Zendy; Canxin Meng | Zendy; Guanpeng Xu | Zendy; Huahua Jian | Zendy; Fengping Wang | Zendy

Open Access

Validation of reference genes for reverse transcription real-time quantitative PCR analysis in the deep-sea bacterium Shewanella psychrophila WP2

Author(s) -

Shunzhang Liu,

Canxin Meng,

Guanpeng Xu,

Huahua Jian,

Fengping Wang

Publication year - 2018

Publication title -

fems microbiology letters

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.899

H-Index - 151

eISSN - 1574-6968

pISSN - 0378-1097

DOI - 10.1093/femsle/fny048

Subject(s) - reference genes , gene , biology , bacteria , shewanella , real time polymerase chain reaction , reverse transcription polymerase chain reaction , gene expression , 16s ribosomal rna , genetics , microbiology and biotechnology

Reference genes are critical to obtain reliable results of reverse transcription real-time quantitative PCR (RT-qPCR), which is widely used for relative quantification of gene expression. In this study, we evaluated the validity of seven candidate reference genes for normalization in RT-qPCR analysis in the deep-sea bacterium Shewanella psychrophila WP2 under different environmental conditions. Among the set of genes investigated, gyrA, 16S rRNA and rho were identified as the most suitable reference genes for WP2 at different temperatures, hydrostatic pressures and salinities, respectively. Notably, the rho gene is conserved in Shewanella genus and other deep-sea bacteria, thus, could be used as a versatile reference gene for RT-qPCR analysis of these microorganisms under extreme environmental conditions.

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