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PmMGST3, a novel microsomal glutathione S-transferase gene in the dinoflagellate Prorocentrum minimum, is a potential biomarker of oxidative stress
Authors:Ruoyu Guo  Vinitha EbenezerJang-Seu Ki
Affiliation:Department of Life Science, Sangmyung University, Seoul 110-743, Republic of Korea
Abstract:In this study, we evaluated a novel microsomal glutathione S-transferase3 (MGST3) gene from the dinoflagellate Prorocentrum minimum, and examined its expression pattern in response to copper-and nickel-induced stresses. The full length of PmMGST3 was 732 bp, ranging from the dinoflagellate splice leader (DinoSL) sequence to the poly (A) tail, covering a 441-bp ORF, 97-bp 5′UTR, and 194-bp 3′UTR. The PmMGST3 was up-regulated by metals, including copper and nickel. The highest up-regulation levels of the PmMGST3 were found under 0.1 mg/L copper and 0.5 mg/L nickel treatment, respectively. In addition, the PmMGST3 was gradually up-regulated by 0.1 mg/L copper with increasing exposure time. Furthermore, ROS production and reduced GSH was measured in the copper treated cells. A significant increased ROS production and reduced GSH were found in the copper treated cells. These results suggest that PmMGST3 may be related to defense mechanisms associated with oxidative stress in dinoflagellates.
Keywords:BLAST, Basic Local Alignment Search Tool   BP, bootstrap proportions   cDNA, complementary DNA   CT, cycle threshold   DHR123, dihydrorhodamine 123   DinoSL, dinoflagellates splice-leader   EC50, median effective concentration   EST, expression sequence tag   GSH, glutathione   GST, glutathione S-transferase   MAPEG, membrane associated protein in eicosanoid and glutathione metabolism   MGSTs, microsomal GSTs   ORF, open reading frame   NJ, neighbor-joining   posterior probabilities, PP   qRT-PCR, quantitative real time polymerase chain reaction   RACE, rapid amplification of cDNA ends   R123, rhodamine 123   ROS, reactive oxygen species   TCA, trichloroacetic acid   TUA, α-tubulin   UTR, untranslated region
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