Cytochrome Oxidase 2 (COX2), β-Tubulin (TUB) and Chitin Synthase Subunit 2 (CHS2) Expression in Pythium insidiosum Thai Strains

Patcharee  KAMMARNJASSADAKUL; Watcharin  RANGSIPANURATN; Methee  SRIPRAPUN; Popchai  NGAMSAKULRUNGRUJ; Tanapat  PALAGA; Kallaya  SRITUNYALUCKSANA; Ariya  CHINDAMPORN

doi:10.48048/wjst.2021.9433

Authors

Patcharee KAMMARNJASSADAKUL Faculty of Medical Technology, Huachiew Chalermprakiat University, Samut Prakan 10540, Thailand
Watcharin RANGSIPANURATN Faculty of Medical Technology, Huachiew Chalermprakiat University, Samut Prakan 10540, Thailand
Methee SRIPRAPUN Faculty of Pharmacy, Mahidol University, Bangkok 10400, Thailand
Popchai NGAMSAKULRUNGRUJ Faculty of Medicine Siriraj Hospital, Mahihol University, Bangkok 10400, Thailand
Tanapat PALAGA Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand
Kallaya SRITUNYALUCKSANA Faculty of Science, Mahidol University, Bangkok 10400, Thailand
Ariya CHINDAMPORN Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand

DOI:

https://doi.org/10.48048/wjst.2021.9433

Keywords:

Chitin synthase 2, Cytochrome oxidase 2, Expression, Pythium insidiosum, Tubulin

Abstract

This study aimed to investigate the real-time fluctuation of temperature-sensitive gene expression in a Pythium insidiosum growing at human body temperature (37 ºC) when it can cause life-threatening disease, whereas its natural habitat is in agricultural water sources with lower temperature. Genes encoding cytochrome oxidase 2 (COX2), β-tubulin (TUB), and chitin synthase subunit 2 (CHS2) were tested for their expression at the infection temperature (37 ºC) and natural habitat temperature (27 ºC) using real-time RT-PCR. Sixteen strains of P. insidiosum, representing 3 phylogeographic preferences, were analyzed. The subculturing process at 27 ºC was repeated at least 2 - 3 cycles. The results showed that at 37 ºC, the COX2 was expressed 2.5-fold higher than that of 27 ºC (p = 0.0347) while the level of TUB and CHS2 mRNA disclosed indistinguishable profiles. These results suggested that these genes were suitable to be used as housekeeping genes for temperature-susceptible gene expression studies. This work was likely to be the 1^st study examining the gene expression levels of the Thai strain of P. insidiosum under thermal stress conditions. Follow-up studies for the COX2 genes may useful to provide valuable insight into the pathogenesis, diagnostic, or therapeutic targets for further investigation.

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