• Title/Summary/Keyword: PAT assay

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Evaluation of the acute toxicity of theoredoxin (TRX) transgenic soybean to Daphnia magna

  • Oh, Sung-Dug;Min, Seok-Ki;Kim, Jae Kwang;Park, Jung-Ho;Kim, Chang-Gi;Park, Soo Yun
    • Korean Journal of Agricultural Science
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    • v.47 no.4
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    • pp.791-802
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    • 2020
  • Theoredoxin (TRX) transgenic soybeans were developed using the human Theoredoxin gene under the control of the ��-conglycinin promoter with a selection marker, the phosphinothricin acetyltransferase (PAT) gene. This study was done to assess the acute toxicity of a genetically modified (GM) soybean using the fresh water planktonic crustacean Daphnia magna. The acute toxicity effect of the TRX soybean and non-GM soybean (Gwangan) on D. magna was investigated at different concentrations (0, 156, 313, 625, 1,250, 2,500, and 5,000 mg·L-1). The TRX soybean used for the test was confirmed to express the TRX/PAT genes by PCR and enzyme-linked immunosorbent assay (ELISA). D. magna feeding tests showed no significant differences in the cumulative immobility or an abnormal response with either the TRX soybean or non-GM soybean. The feeding study showed a similar abnormal response and cumulative immobility of the D. magna between the TRX soybean and Gwangan treatments. Additionally, the 48 h-EC50 values for the TRX and Gwangan soybeans were 755.6 and 778 mg·L-1, respectively. The soybean NOEC (no observed effect concentration) value for D. magna was suggested to be 156 mg·L-1. These results suggest that there is no significant difference in toxicity to Daphnia magna between the TRX soybean and its non-GM counterpart.

Alaternin and Emodin with Hydroxyl Radical inhibitory and/or Scavenging Activities and Hepatoprotective Activity on Tacrine-Induced Cytotoxicity in HepG2 Cells

  • Jung, Hyun-Ah;Chung, Hae-Young;Takaka, Yokezawa;Kim, Youn-Chul;Hyun, Sook-Kyung;Choi, Jae-Sue
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.947-953
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    • 2004
  • The antioxidative and hepatoprotective potentials of two anthraquinones, alaternin (2-hydroxy-emodin) and emodin, to scavenge and/or inhibit hydroxyl radicals generated by the Fenton reaction and to protect tacrine-induced cytotoxicity in human liver derived HepG2 cells were evaluated, respectively. The inhibitory activity on hydroxyl radical generated in a cell-free chemical system (FeSO$_4$/$H_2O$$_2$) was investigated by a fluorescence spectrophotometer using a highly fluorescent probe, 2$^1$,7$^1$-dichlorofluorescein. The hydroxyl radical scavenging activity was determined by electron spin resonance spectroscopy using 5,5-dimethy-1-pyrroline-N-oxide as hydroxyl radicals trapping agents. Tacrine-induced HepG2 cell toxicity was determined by a 3-[4,5-dimethylthiazole-2yl]-2,5-diphenyltertrazolium bromide assay. Although the scavenging activity of alaternin on hydroxyl radical was similar to that of emodin in dose-dependent pat-terns, the inhibitory activity exhibited by the former on hydroxyl radical generation was stron-ger than that of the latter, with $IC_{50}$/ values of 3.05$\pm$0.26 $\mu$M and 13.29$\pm$3.20 $\mu$M, respectively. In addition, the two anthraquinones, alaternin and emodin showed their hepatoprotective activ-ities on tacrine-induced cytotoxicity, and the EC$_{50}$ values were 4.02 11M and 2.37 $\mu$M, respec-tively. Silymarin, an antihepatotoxic agent used as a positive control exhibited the EC$_{50}$ value of 2.00 $\mu$M. These results demonstrated that both alaternin and emodin had the simultaneous antioxidant and hepatoprotective activities.ies.

Influence of heading date difference on gene flow from GM to non-GM rices (GM벼에서 non-GM벼로 유전자 이동에 대한 개화기 차이의 영향 분석)

  • Oh, Sung-Dug;Chang, Ancheol;Kim, Boeun;Sohn, Soo-In;Yun, Doh-Won
    • Journal of the Korean Society of International Agriculture
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    • v.30 no.4
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    • pp.347-356
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    • 2018
  • Genetically modified (GM) crops have been increased continuously over the world and concerns about the potential risks of GM crops have also been increasing. Even though GM crops have not been cultivated commercially in Korea, it should be necessary to develop the safety assesment technology for GM crops. In this study, we investigated the influence of heading date difference on gene flow from GM to non-GM rice. In the experimental plot design, The PAC GM rice was placed in the center as a pollen donor and non-GM rice were placed in eight directions as pollen receivers. Five pollen receiver rice cultivars were Unkawng, Daebo, Saegyejinmi, Nakdong-byeo, and Ilmi which had different flowering times. A total of 266,436, 300,237, 305,223, 273,373, and 290,759 seeds were collected from Unkawng, Daebo, Saegyejinmi, Nakdong, and Ilmi, respectively, which were planted around PAC GM rice. The GM${\times}$non-GM hybrids were detected by repeated spraying of herbicide and PAT immunostrip assay. Finally, the hybrids were confirmed by PCR analysis using PAC gene specific primer. The hybrids were found in Nakdong-byeo which had the same heading date with PAC GM rice. The hybridization rate was 0.0007% at Nakdong-byeo plot. All of GM${\times}$non-GM hybrids were located within 2 m distance from the PAC GM rice zone. The physiological elements including rice heading date were found to be important factors to determine GM?rice out crossing rate with GM rice. Consideration should be taken into for many factors like the physiological elements of field heading date of rice cultivars to set up the safety management guideline for prevention of GM rice gene flow.