Low-level lasers and mRNA levels of reference genes used in Escherichia coli. (13th October 2016)

• Record Type: Journal Article
• Authors: Teixeira, A F; Machado, Y L R C; Fonseca, A S; Mencalha, A L
• Is Part Of: Laser physics letters. Volume 13:Number 11(2016:Nov.)
• Publication Date: 2016-10-13
• Subjects: low-level laser therapy -- gene expression -- RT-qPCR -- housekeeping; Lasers -- Periodicals; Physics -- Periodicals; 621.366
• DOI: 10.1088/1612-2011/13/11/115602
• Languages: English
• ISSNs: 1612-2011

Abstract

Abstract: Low-level lasers are widely used for the treatment of diseases and antimicrobial photodynamic therapy. Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) is widely used to evaluate mRNA levels and output data from a target gene are commonly relative to a reference mRNA that cannot vary according to treatment. In this study, the level of reference genes from Escherichia coli exposed to red or infrared lasers at different fluences was evaluated. E. coli AB1157 cultures were exposed to red (660 nm) and infrared (808 nm) lasers, incubated (20 min, 37 °C), the total RNA was extracted, and cDNA synthesis was performed to evaluate mRNA levels from arcA, gyrA and rpoA genes by RT-qPCR. Melting curves and agarose gel electrophoresis were carried out to evaluate specific amplification. Data were analyzed by geNorm, NormFinder and BestKeeper. The melting curve and agarose gel electrophoresis showed specific amplification. Although mRNA levels from arcA, gyrA or rpoA genes presented no significant variations trough a traditional statistical analysis, Excel-based tools revealed that these reference genes are not suitable for E. coli cultures exposed to lasers. Our data showed that exposure to low-level red and infrared lasers at different fluences alter the mRNA levels from arcA, gyrA and rpoA in E. coli cells.