Glutathione S-transferase (GST) is a key gene involved in multiple stress tolerance in all living organisms, though it is still to be disclosed the gene function in teff grass [Eragrostis tef (Zucc.)Trotter].The objectives of this study were to clone and molecular characterization of GST gene in teff grass. We characterized GST1 from teff grass (EtGST1), it composed of a 645-bp open reading frame (ORF) that encoded 195 amino acid residue. Further, we transformed EtGST1 in E.coli BL21 (DE3) cells. This recombinant EtGST1 in E.coli BL21(DE3) induced at 37°C temperature. In addition, Growth of cells overexpressing EtGST1 rapidly increased in the presence of polyethylene glycol (5%), heat (46°C), NaCl (0.6%), and arsenic (1 mM) than that of cells harboring an empty vector. These results suggest that EtGST1 would be suitable candidate for improving tolerance in forages and/or grasses species against multiple abiotic stresses.

카드뮴과 같은 중금속은 독성이 높아 수서 생물과 인간에게 해로운 영향을 미친다고 알려져 있다. 본 연구에서는 해양 섬모충 Euplotes crassus에서 카드뮴이 해독 기전에 관여하는 ABC transporters (ABCs)와 glutathione S-transferase (GST)의 유전자 발현에 미치는 영향을 조사하였 다. 총 7개의 ABCs 유전자와 1개의 GST 유전자 일부를 클로닝하여 유전자 분석을 실시하였고, 카드뮴(0.1~1 mg/l) 노출에 따른 이들 유전자의 발현 양상을 quantitative real time RT-PCR (qRT-PCR)을 이용하여 분석하였다. 염기서열 분석과 계통 분석 결과 이들 ABCs 유전자가 ABC transporter의 특징을 가지며, ABC-B/C family에 속하는 것을 확인하였고, GST 유전자는 theta isoform과 유사한 것으로 나타났다. 카드뮴에 8시간 노출시킨 결과 ABC transporter 유전자의 경우 ABCB21 유전자를 제외하고는 대부분 농도 의존적으로 유전자 발현이 유의하게 증가하였 다. GST 유전자는 0.5 mg/l에서 가장 높은 유전자 발현 양상을 보였으며, 1 mg/l에서는 발현량 이 대조군 수준으로 감소되었다. 본 연구 결과는 E. crassus의 ABC transporter와 GST 유전자가 카드뮴에 의해 유도되는 독성에 대한 방어 기전에 참여하는 것을 의미한다.

Vitellogenins (Vgs) are precursors of the major egg storage protein, vitellin (Vn), in many oviparous animals. Insects Vgs are large molecules (200-kD) synthesized in the fat body in a process that involves substantial structural modifications (e.g., glycosylation, lipidation, phosphorylation, and proteolytic cleavage, etc.) of the nascent protein prior to its secretion and transport to the ovaries. However, the extent to which Vgs are processed in the fat body varies greatly among different insect groups. We were cloned Vgs partial genes PaVgs and BgVgs from Periplaneta americana and Blattella germanica. Real-time quantitative PCR shows that PaVgs and BgVgs were differential-regulated with aging. In insects, glutathione S-transferases (GSTs) are enzymes involved in detoxification of insecticides. We were cloned GST partial genes PaGST and BgGST from Periplaneta americana and Blattella germanica. Real-time quantitative PCR shows that PaGST and BgGST were up-regulated with aging, and the mRNA level of PaGST and BgGST was higher in 4℃ and 37℃ than room temperature. The expression level of PaGST and BgGST exposure to temperature stress suggests that PaGST and BgGST are up-regulated after exposure low and hige temperature treatments.

본 연구는 시험생물로서 말똥성게 (Hemicentrotus pulcherrimus) 를 이용하여 내분비계장애물질인 bisphenol A (BPA)의 독성 및 시험생물로서의 적합성 등을 조사하였 다. 말똥성게(H. pulcherrimus)의 수정 및 정상 배아발생 에 미치는 BPA의 독성을 보기 위하여 농도(0, 300, 500, 800, 1000, 1500 ppb)에서 조사하였다. BPA 노출 시 수정 률은 시험구간 내의 BPA 처리농도와 관계없이 유의적인 변화가 없었다. 정상 배아발생률은 BPA 농도가 높을수 록 유의적인 감소를 나타냈으며, 800 ppb 농도부터 유의 적인 감소를 보였다. 정상배아발생에 대한 독성값은 반 수영향농도 (EC50) 1056.1 ppb, 95% Cl 981.8~1163.9 ppb 로 나타났다. 또한 무영향농도 (NOEC)와 최소영향농도 (LOEC)는 500 ppb 및 800 ppb로 나타났다. BPA에 노출 된 배아는 농도가 증가함에 따라 발생이 정체되는 현상 이 나타났다. BPA에 노출된 pluteus 유생을 이용한 glutathione- S-transferase (GST) 유전자의 발현을 비교해본 결과 GST 유전자의 발현은 농도가 증가함에 따라 발현 이 증가하였다. 본 연구 결과, 말똥성게 (H. pulcherrimus)의 초기 배아 발생 과정 중 800 ppb 이상에서 독성을 나타냈으며 GST 유전자는 BPA 노출에 따른 위해성 평가에 생체지표유 전자로 유용하게 이용될 수 있다고 생각된다.

70% 에탄올로 양파를 추출한 다음 감압하에 농축하였고, 농축된 양파 추출물을 증류수에 재 용해시킨 용액의 pH는 5.34인 약산성으로 나타났다. 그리고 이 용액을 0.1 N NaOH 로 적정한 다음 TLC(전개용매: ethyl acetate)에서의 분리는 적 정 전보다 chromatogram의 이동 및 분리가 뛰어난 것으로 나 타났고, GST 활성 저해에도 유의성이 있는 것으로 나타났다. Silica-gel column chromatography에서 0.1 N NaOH 적정용액의 분리는 ethyl acetate(분획 A), methanol(분획 B), 50% methanol (분획 C)로 용출하였고, 용출된 각 분획은 감압하에 농축하여 GST 활성 저해 실험을 하였다. 세 부분 모두 GST 활성 저해에 유의성이 있는 것으로 나타났다. 각 분획은 GC-MS spectrometer 분석에서 두 종류 이상이 함유되어 있지만, 구성 물질은 서로 상이함을 보였고, 이들 물질의 분리와 구조 규명이 필요한 것 으로 사료된다.

Glutathione S-transferases (GSTs) are multifunctional enzymes that are mainlyinvolved in the xenobiotic metabolism and protection against oxidative damage. Most studies of GSTs in insects have been focused on their role in detoxifying exogenous compounds in particular insecticides. Here, we show the expression profiles of GSTs of the bumblebee Bombus ignitus in response to oxidative stress. We identified a sigma-class GST from B. ignitus (BiGSTS). The BiGSTSgene consists of 4 exons that encode 201 amino acids. Comparative analysis indicates that the predicted amino acid sequence of BiGSTS shares a high identity with the sigma-class GSTs of hymenopteran insects such as Apis mellifera (70% protein sequence identity) and Solenopsis invicta (59% protein sequence identity). Tissue distribution analyses showed the presence of BiGSTS in all tissues examined, including the fat body, midgut, muscle and epidermis. The oxidative stress responses analyzed by quantitative real-time PCR showed that under H2O2 overload, BiGSTS and BiGSTD (identified in our previous study) were upregulated in all tissues examined, including the fat body and midgut of B. ignitus worker bees. Under uniform conditions of H2O2 overload, the expression profile of GSTs and other antioxidant enzyme genes, such as phospholipid-hydroperoxide glutathione peroxidase (Bi-PHGPx) and peroxiredoxins (BiPrx1 and BiTPx1), showed that other antioxidant enzyme genes are acutely induced at 3 h after H2O2 exposure, whereas BiGSTS and BiGSTD are highly induced at 9 h after H2O2 exposure in the fat body of B. ignitus worker bees. These findings indicate that GSTs and other antioxidant enzyme genes in B. ignitusare differentially expressed in response to oxidative stress. Taken together, our findings indicate that BiGSTS and BiGSTD are oxidative stress-inducible antioxidant enzymes that may play a role in oxidative stress response.

The effects of different dietary fatty acids on the hepatic glutathione S-transferase(GST-P) positive foci and glutathione related enzyme system were investigated in carcinogen treated rats. Weaning male Sprague Dawley rats were divided into three groups and fed the diets of 15% corn(CO), perilla(PO), and sardine oil(SO), respectively. Hepatocellular carcinogenesis was initiated with diethylnitrosamine(DEN) and then fed the diet containing 0.02% 2-acetylaminoflumene(2-AAF) followed by 0.05% phenobarbital for 10 weeks. The hepatic tissues were homogenized and centrifugated to prepare microsoma1 and cytosolic fractions. The enzyme activities of hepatic glutathione S-transferase(GST), glutathione reductase(GR), and glutathione peroxidase(GPx) were determined from cytosoIic Mans. The number of GST-P hyperplastic nodules was the highest in corn oil group at 6th week, the early stage of hyperplastic nodule formation. GST activities were increased significantly by carcinogens in a11 dietary groups after 6th wk. GR activities followed the same tread as GST activities. GPx activities were decreased by carcinogens in all dietary groups at 10th week. In this experiment, corn oil diet may have promotive effect on hyperplastic nodule formation during the early promotional stages of chemical carcinogenesis.

방오도료로 많이 쓰이던 유기주석화합물은 일반생물에게 미치는 독성이 매우 강하고 또한 내분비계 장애물질임이 밝혀지면서 이를 대체할 화합물들이 개발되고 있다. 그 가운데 본 연구에서는 Sea-Nine 211(4,5-dichlore-2-n-octyl-3(2H) isothiazolone)을 사용하여 이 화합물이 해양생물 특히 저서생물인 패류에게 얼마나 영향을 미치는지를 살펴보았다. 이를 위해 강원도 북부 해역에 주로 서식하는 북방대합(Pseudocardium

This study was performed to investigate antioxidant activities and glutathione S-transferase (GST) activity according to parts of the Acer mono and A. okamotoanum. Most extracts showed high scavenging activities on DPPH. Especially, the bark of A. okamotoanum showed higher activity as 98.4% than the control, BHA as 96.5%. A. mono and A. okamotoanum showed high ability on nitrite scavenging, but decreasing tendency according to decreasing of pH. On SOD-like test, the wood of A. okamotoanum showed highest activity as 35.4% at 1.0mg/ml concentration. Also, the extracts obtained high activity on GST test. Therefore, the water extracts from the bark of A. mono and A. okamotoanum have relatively good antioxidant activity and GST activity. Especially, the bark of A. okamotoanum showed the highest activity on all of extracts, could be the use of functional foods and biomaterials.

더덕 뿌리에서 유래한 EST cDNA library로부터 GST(glutatione S-transferase)유전자와 높은 상동성을 나타내는 full clone cDNA를 얻었다. 더덕의 GST(glutatione S-transferase), CIGST은 761 bp의 cDNA로 173개의 아미노산을 코딩하는 522 bp의 ORF를 가지고 있으며, A. thaliana(AAC63629) 71%, C. chinense(CAI51314) 73%, E. esula(AAE65767) 75%, H. muticus(CAA55039) 70%, N. plumbaginifolia(CAA96431) 77%, S. commersonii(AAB65163) 76%등 다른 식에서 밝혀져 있는 GST(glutatione S-transferase)와 유의한 상동성을 나타내였다.

Using Agrobacterium-me야ated transformation method the auxin-regulated cotton GST (Gh-5) constructs were used to transform Rehmannia glutinosa L. The PCR analysis was conducted to verify transgenicity. Based on the PCR analysis, there was verified that the 988 bp DNA band had showed in transgenic plant genomes in PCR anaJysis using Gh5-1 and Gh5-2 primers. The effects of cocultivation with Agrobacterium tumefaciens, regeneration and selection conditions on the transformation efficiency of Chinese foxglove (Rehmannia glutinosa L.) were investigated. Factors such as cocultivation period, use of acetosyringone, postcultivation in darkness, and different kanamycin concentrations for selection were assessed. In vitro regeneration, the number of leaves, shoot lengths and numbers on MS medium were superior to on B5 and WPM medium, and the shoot formation rate was highest level of 95% in cultured base part containing leaf stalk. Addition of acetosyringone at concentration of 200μM to cocultivation medium and 3-day of cocultivation improved transformation frequencies. Exposure of explants to darkness for 4 weeks on selection medium resulted in further increased the regeneration frequency of transgenic shoots. In PCR analysis, the amplified fragments of Gh5 gene were detected (988 bp), and GST-expressing transgenic R. glutinosa L. plants had approximately three-fold higher activity in leaf extracts compared with control plant.

A gene coding for the GST of cotton (Gh-5) was cloned into Escherichia coli and experssed. The enzyme remained within the cytoplasm of E. coli. An 696 bp open reading frame was in the 988 base pair fragment of the recombinant plasmid pET-30b(+). The deduced protein sequence consists of 232 amino acids and has a molecular mass of 30235.58 Da. The cloned enzyme conjugated reduced glutathione and 1-chloro-2,4-dinitrobenzene (CDNB). Plant GST cDNA was expressed in microbe and produced polypeptide had function as an enzyme.