杀虫单对二化螟谷胱甘肽转移酶(GSTs)的抑制作用

以二化螟为材料研究了杀虫单、甲胺磷和顺丁烯二酸二乙酯 (DEM)对谷胱甘肽转移酶 (GSTs)的抑制作用。离体抑制试验结果表明 ,杀虫单和DEM都对二化螟GSTs活性有强烈的抑制作用 ,在实验设置的反应体系中 ,2 5mmol·L-1杀虫单和DEM与酶液保温 2min后 ,对GSTs的抑制率达到 70 %以上 ;6min后 ,达到 90 %以上。杀虫单和DEM的抑制中浓度 (IC50 )分别是 1 33和 1 71mmol·L-1,而甲胺磷没有抑制作用。活体实验结果显示 ,用 0 1mol·L-1杀虫单、甲胺磷和DEM 0 0 5 μL分别处理二化螟 4龄幼虫 48h后 ,杀虫单和DEM对GSTs活性的抑制率分别达 10 3%和 14 5 % ,甲胺磷则表现出 8 6 %的诱导作用。酶动力学实验显示 ,杀虫单和DEM都是GSTs的可逆性抑制剂 ,这种抑制作用不是竞争性的。由此表明 ,杀虫单是GSTs的强抑制剂 ,这在杀虫单的毒杀机制中具有重要的意义。

花生GSTs家族基因的全基因组分析

谷胱甘肽转移酶(glutathione-S-transferase,GST)是一类重要的多功能超家族酶,普遍存在于植物体中,作为配体或结合蛋白,在调控植物的生长发育、解除外界毒素作用过程起到重要作用。为全面了解花生GSTs基因家族的结构特性,本论文通过生物信息分析技术对花生GST基因家族进行全基因组鉴定和表达特性分析,结果表明,在花生全基因组中一共有163个GSTs基因,其中A组基因中含有76个,B组基因中含有87个。基因结构和发育进化树将花生GST基因分为6类:Tau、Theta、Lamda、EF1Bγ、Phi和DHAR,花生GST家族基因结构进化相对保守,但各基因的内含子数量及长度有较大差异,其中EK5R9的外显子数最多,达19个;而K7JNV的内含子最长,约18 kb。MEME分析发现,花生GST家族基因中存在9个保守域,各基因中的保守结构域为6~8个不等。染色体定位显示,花生GSTs家族基因在花生A、B染色体组上呈不均匀分布,在A02、A03、A07、A09、B02、B03、B05、B08和B09号染色体上存在1~2个基因簇。表达分析发现,只有80个花生GST基因具有组织表达差异,其中6个的组织表达有极显著差异。本研究为GSTs基因的功能研究及花生抗逆性提高提供了理论依据。

溴氰菊酯对麦穗鱼谷胱甘肽S-转移酶(GSTs)的影响

研究了溴氰菊酯不同浓度、不同处理时间对麦穗鱼体内谷胱甘肽S-转移酶(GSTs)活性的影响,同时测定了麦穗鱼不同组织中谷胱甘肽S-转移酶对低浓度溴氰菊酯处理的反应特点。结果表明,处理48 h后,高浓度(0.009 mg/L)组对肝中GSTs活性的抑制率达到43.7%,对鳃中GSTs活性的抑制率达54.3%;而低浓度组(0.001 mg/L)对鳃中GSTs活性有一定的诱导作用。亚致死剂量的溴氰菊酯(3.0×10-4mg/L)处理96 h后,对卵巢的诱导活性最高,其GSTs活性是对照的2.43倍,但对肝、鳃、肾、肠的诱导作用均较低。溴氰菊酯对麦穗鱼不同组织GSTs活性诱导的时间效应和强度不同。

Polymorphisms of the oxidant enzymes glutathione S-transferase and glutathione reductase and their association with resistance of Plasmodium falciparum isolates to antimalarial drugs

Objective:To investigate the association between amplification of the two regulatory genes controlling glutathione(GSH) levels,glutathione reductase(PfGR) and glutathione S-transferase (PfGST) genes and sensitivity of Plasmodium falciparum(P.falciparum) isolates collected from different malaria endemic areas of Thailand to standard antimalarial drugs.Methods:A total of 70 P.falciparum isolates were collected from endemic areas of multi-drug resistance (Tak,Chantaburi and Ranong Provinces) during the year 2008-2009.The in vitro assessment of antimalarial activity of P.falciparum clones(K1- and Dd2 chloroquine resistant and 3D7- chloroquine sensitive) and isolates to chloroquine,quinine,mefloquine and arteusnate was performed based on SYBR Green modified assay.Results:68(97.14%),11(15.71%) and 28(40%) isolates respectively were classified as chloroquine-,quinine- and mefloquine-resistant isolates. With this limited number of P.falciparum isolates included in the analysis,no significant association between amplification of PfGST gene and sensitivity of the parasite to chloroquine, quinine,mefloquine and quinine was found.Based on PCR analysis,Dd2,Kl and 3D7 clones all contained only one copy of the PfGST gene.All isolates(70) also carried only one copy number of PfGST gene.There appears to be an association between amplification of PfGR gene and chloroquine resistance.The 3D7 and Dd2 clones were found to carry only one PfGR gene copy, whereas the K1 clone carried two gene copies.Conclusions:Chloroquine resistance is likely to be a consequence of multi-factors and enzymes in the GSH system may be partly involved. Larger number of parasite isolates are required to increase power of the hypothesis testing in order to confirm the involvement of both genes as well as other genes implicated in glutathione metabolism in conferring chloroquine resistance.

Cloning, characterization and expression analysis of a microsomal glutathione S-transferase gene from the seagrass Zostera marina

The response of glutathione S-transferase(GST)in Zostera marina to temperature variation was analyzed at molecular level by cloning the microsomal GST gene and texting the microsomal GST expression regularity under different temperature.Specific speaking,express ZmGST in Escherichia coli,then purify the recombinant protein and make the thermal stability analysis.Therefore,the experiments were carried out to provide a theoretical basis for the further elaboration to the population degradation mechanisms of Z.marina.In conclusion,the thermostability and the response of ZmGST gene to temperature changes can determine its temperature tolerance range,and affect its resilience in turn.

Comparative Studies of Substrate and Inhibitor Specificity of Glutathione S-Transferases in Six Tissues of Oxya chinensis(Thunberg)(Orthoptera:Acrididae)

Specific activity, substrate specificity, and kinetic parameters （Km and Vmax） of glutathione S-transferases （GSTs） towards three substrates, 1-chloro-2,4-dinitrobenzene （CDNB）, 1,2-dichloro-4-nitrobenzene （DCNB）, and p-nitrobenzene chloride （pNBC） were investigated in six tissues （foregut, midgut, hindgut, fat body, hemolymph, and muscle） of Oxya chinensis. In addition, the inhibition in vitro （ethacrynic acid, and Cibacron Blue 3GA） of Oxya chinensis in the six tissues was also investigated. Glutathione S-transferase activity was detected in all the six tissues examined. The rank order of GST activities towards CDNB was fat body 〉 midgut 〉 hindgut 〉 muscle 〉 foregut 〉 hemolymph both in females and males. Glutathione S-transferase activities in the fat body in females and males were 1.3- to 10.4-fold and 1.1- to 10.0- fold higher than those in the other tissues. The rank order of GST activities towards the other substrates changed slightly. From these results, it was inferred that GSTs in the fat body and midgut played important roles in detoxifying xenobiotics including insecticides and plant allelochemicals in O. chinensis. In the three substrates examined, CDNB seemed to be the best substrate, followed by pNBC and DCNB. The kinetic parameters of GSTs were different among the six tissues. This suggested that GSTs in different tissues have various affinities and catalytic efficiency to substrates. In vitro inhibition study showed that the median inhibition concentration （IC50） values of the two inhibitors to GSTs from the six tissues were different. The results suggested that the two inhibitors have different inhibition potency to GSTs from the different tissues. The observed changes in kinetic parameters and inhibition in vitro among the six tissues of the insect might suggest that the number and structure of isoenzymes and their rate of expression varied for the different tissues.

Antioxidant Effect of Selenium-containing Glutathione S-Transferase in Rat Cardiomyocytes

As one of the most important antioxidant enzymes, glutathione peroxidase（GPX） protects cells and tissues from oxidative damage, and plays an important role in cardiovascular and cerebrovascular injuries induced by oxida- tive stress. The antioxidant effect of selenium-containing glutathione S-transferase（Se-GST）, a mimic of GPX was investigated on rat cardiomyocytes. To explore the protection function of Se-GST in hydrogen peroxide（H202） chal- lenged rat cardiomyocytes, we examined malondialdehyde（MDA）, lactate dehydrogenase（LDH）, superoxide dismu- tase（SOD） and cell apoptosis. The results demonstrate exposure of rat cardiomyocytes to H202 for 6 and 12 h induced the significant increases of MDA, LDH and apoptosis rate of cardiomyocytes, but pretreatment of rat cardiomyocytes with Se-GST at 0.0005 or 0.001 unit/mL prevents oxidative stress induced by H202 with the decreases of cell apopto- sis. All the results hint Se-GST has antioxidant activity for oxidative stress challenged rat cardiomyocvtes.

星豹蛛两个GST基因克隆、鉴定及其对溴氰菊酯胁迫的响应表达

【目的】为明确星豹蛛Pardosa astrigera体内2个谷胱甘肽S-转移酶(glutathione S-transferases,GSTs)基因的时空表达模式,并探究其是否能对溴氰菊酯的胁迫做出响应。【方法】基于星豹蛛转录组数据库,PCR克隆星豹蛛GST基因PaGSTd1和PaGSTd2全长cDNA序列并进行生物信息学分析;利用RT-qPCR检测PaGSTd1和PaGSTd2在星豹蛛不同发育阶段(2-6龄若蛛和成蛛)、雌雄成蛛不同组织(头胸部、腹和足)以及通过药膜法利用不同浓度[LC 10(5.151 mg/L),LC 30(8.619 mg/L)和LC 50(12.311 mg/L)]溴氰菊酯胁迫不同时间(6,12,18,24和48 h)雄成蛛中的表达量。【结果】星豹蛛PaGSTd1(GenBank登录号:OR096398)全长cDNA序列长708 bp,开放阅读框长645 bp,编码214个氨基酸;星豹蛛PaGSTd2(GenBank登录号:OR096399)全长cDNA序列长793 bp,开放阅读框长645 bp,编码214个氨基酸。RT-qPCR检测结果表明,PaGSTd1和PaGSTd2在星豹蛛不同发育阶段和成蛛不同组织都有表达,均在5龄若蛛中的表达量最低;PaGSTd1在4龄若蛛中的表达量最高,PaGSTd2在成蛛中的表达量最高;PaGSTd1和PaGSTd2在足中的表达量最高,在除腹部外的其他组织中表达量均为雄成蛛的显著高于雌成蛛的。LC 10溴氰菊酯胁迫24 h时星豹蛛雄成蛛中PaGSTd1被诱导表达,6和18 h时PaGSTd2被诱导表达;LC 30溴氰菊酯胁迫18,24和48 h时雄成蛛中PaGSTd1被诱导表达,18 h时PaGSTd2被诱导表达;LC 50溴氰菊酯胁迫24 h时雄成蛛中PaGSTd1和PaGSTd2被诱导表达。【结论】本研究克隆得到星豹蛛两个GST基因PaGSTd1和PaGSTd2,均在星豹蛛足中表达量最高,说明足中这2个GST基因在对外源物质的解毒起到重要作用;这两个GST基因可以被溴氰菊酯诱导表达,说明可能参与星豹蛛对溴氰菊酯的胁迫响应,为后续GSTs的功能研究提供线索。

Expression and Characterization of a Sigma-Class Glutathione S-transferase of the Oriental Migratory Locust, Locusta migratoria manilensis (Meyen)

A cDNA encoding a sigma-class glutathione S-transferase of the locust, Locusta migratoria manilensis （LmGSTs1）, was cloned by reverse transcriptase-polymerase chain reaction. The 830 bp-long cDNA encoded a 615 bp open reading frame （204 amino acid polypeptide）, which exhibited the structural motif and domain organization characteristic of GST sigma-class. It revealed 59, 57, 57, and 56% identities to sigma-class GSTs from Blattella germanica, Gryllotalpa orientalis, Nasonia vitripennis, and Pediculus humanus corporis, respectively. A recombinant protein （LmGSTs1） was functionally expressed in Escherichia coli cells in a soluble form and purified to homogeneity. LmGSTs1 was able to catalyze the biotranslation of glutathione with 1-chloro-2,4-dinitrobenzene, a model substrate for GSTs, as well as with p-nitro-benzyl chloride. Its optimal activity was observed at pH 8.0 and at 30℃. Incubation for 30 min at temperatures below 50℃ scarcely affected the activity. The I50 of reactive blue （RB） was 18.5 μmol L-1. In the presence of 0.05 mmol L-1 ethacrynic acid （ECA）, LmGSTs1 showed （81±3）% of the original activities.

Antibodies against ribosomal protein S29(RPS29)fused with glutathione's transferase specially react with native RPS29 in mouse and human cells

The ribosomal protein S29 also known as RPS29, is not only a component of the 40S subunit of ribosome, but also involved in embryonic development, oncogenesis and other pathologic conditions. However, rare commercial antibody against RPS29 restricts the discovery of precise physiological and pathological function of this protein. In this study, the whole RPS29 gene was inserted into plasmid pGEX-6p-1 to express glutathione＇s transferase （GST） fusion proteins in Escherichia eoli （E. coli） strain BL21. High yields of soluble recombinant proteins were obtained. Mice were immunized with the recombinant RPS29 protein. The serum from the immunized mice could specially react with purified recombinant RPS29 proteins and native RPS29 proteins in CCE cells by western blotting, immunofluorescence staining and flow cytometric analysis. Further more the polyclonal antibodies also reacted specifically with human cell strain ECV304, which showed typical cytoplasmatic fluorescence. The polyclonal antibodies we prepared would be an available tool for studying the roles of RPS29 in embryonic development and human diseases.

Frequency of Null Phenotypes of Glutathione S-Transferase M1 and T1 among the Populations of Tabuk (Northwestern Part of Saudi Arabia)

Background: The variability in the distribution of the null phenotypes of GSTM1 and GSTT1, due to total or partial gene deletion resulting in the lack of the active enzyme, has been reported in different populations, especially in ethnically well-defined groups but not in Tabuk. This study investigated the variability in the distribution of the null phenotypes of GSTM1 and GSTT1 in the population of Tabuk (northwestern part of Saudi Arabia). Method: This study was conducted on 200 subjects of Tabuk—northwestern part of Saudi Arabia among which 100 were chronic smokers and 100 were nonsmokers. The subjects were reporting to hospital for routine checkup. All were without past history of any chronic disease and no significant abnormality. GST genotyping was done by multiplex PCR-based methods. The smoker and control groups were compared using a chi-square test with P GSTM1 deletion homozygosity of 14% and 1% was reported among non smokers and smokers, respectively whereas GSTT1 deletion homozygosity of 28% and 6% was reported among non smokers and smokers, respectively. Our results indicate that there are major differences in allelic distribution of GSTM1 and GSTT1 genes between the two groups investigated. Combined analysis of both genes revealed that 15% of smokers and non smokers harbor the deleted genotype of GSTM1 and 34% of smokers and non smokers harbor the deleted genotype of GSTT1 with significant differences. Conclusion: This study enables selecting subgroups among the general population who are more susceptible to DNA damage and will help genetic studies on the association of GST polymorphisms with disease risks and drug effects in Arab population. Studies with a larger sample size are needed to evaluate and confirm the validity of our results.

小麦抗白粉病相关基因GST克隆与表达

以从小麦抗白粉病相关基因差异表达分析中获得的EST-3(Genbank序列号EX567360)为标签,采用电子克隆的方法对其进行延伸,并对电子克隆结果进行半定量RT-PCR验证,最后对白粉菌不同侵染时间进行了表达分析。经RT-PCR扩增,EST-3表达的带型变化趋势与其在抑制性消减杂交SSH-cDNA的差异显示情况一致,且RT-PCR获得的序列与电子克隆的序列一致性达98%。生物信息学分析表明,该序列是由875bp核苷酸组成的,具有完整的开放阅读框架,编码蛋白为229个氨基酸,GenBank序列号JK841279,含有一个N端和C端谷胱甘肽硫转移酶结构域,该序列与小麦谷胱甘肽硫转移酶基因(GST)一致性较高,达97%。表达分析结果显示,白粉菌侵染24h表达受到抑制,48h开始表达,侵染72h表达最强,96h又开始下降,表明GST基因属于白粉菌诱导型相关基因,参与小麦对白粉病的应答反应。

日本血吸虫重组28GST T细胞表位谱的预测及鉴定

目的 :用软件预测日本血吸虫重组 2 8GST抗原分子T细胞表位谱 ,并鉴定其Th1型细胞表位。方法 :用软件预测重组2 8GSTT细胞表位谱 ,并筛选几种较好的T细胞表位 ,人工合成表位肽或用基因工程制备重组表位肽融合蛋白。体外刺激经照射的尾蚴感染并用 2 8GST加强免疫的C5 7BL/ 6小鼠 (H 2 b)脾细胞 ,通过淋巴细胞增殖试验、ELISA及流式细胞术等 ,分析各种表位的免疫刺激作用 ,鉴定Th1型细胞表位。结果 :在 9个候选的表位中 ,P6 (73～ 86aa)是刺激作用最强的Th1型细胞表位。结论 :重组 2

陆地棉GST基因家族全基因组分析

谷胱甘肽转移酶(glutathione-S-transferase,GST)是一种普遍存在的具有多功能的超家族蛋白,在植物初次生代谢、逆境胁迫、胞间信号传递等方面具有重要作用;同时,作为配体其在植物激素代谢以及物质转运方面也发挥作用。为了解析陆地棉(Gossypium hirsutum L.)GST基因家族的信息,本研究对该基因家族成员的种类、进化关系、物理定位、基因结构和保守基序以及表达模式进行了分析。结果显示,在陆地棉全基因组中共含有70个GST基因,进化树和基因结构分析将该家族分为U族、F族、T族、Z族、EF1Bγ族和TCHQD族。基因定位分析发现,除了AD/At2、AD/At4、AD/At5、AD/Dt5、AD/Dt10号染色体上没有GST基因外,其他染色体上都有GST基因,并且在AD/At9、AD/Dt7、AD/Dt12、AD/Dt13这4条染色体上出现基因簇。对F族(Phi类)9个GST基因进行荧光定量分析,结果表明,除Gh GSTF1可能为假基因外,Gh GSTF2~9等8个基因在陆地棉根、茎、叶以及各个发育时期的纤维中均有表达;结合生物信息学分析,推测Gh GSTF8可能参与原花青素/花青素的转运和积累;Gh GSTF4、Gh GSTF6和Gh GSTF9可能在调节陆地棉的生长和胁迫反应中起作用,而Gh GSTF2、Gh GSTF3、Gh GSTF5和Gh GSTF7的功能还有待进一步研究。本研究为陆地棉GST基因家族的分子进化及功能研究提供了理论依据。

GSTM1与GSTP1基因多态性对红细胞GST酶活性的影响

目的研究单独或结合GSTM1纯合缺失基因型时,不同GSTP1基因型对红细胞GST酶活性的影响。方法GST酶活性参照Habig等报道的方法用紫外分光光度法测定。用多重PCR分析GSTM1基因多态性,PCRRFLP检测GSTP15号外显子105位密码子基因多态性。结果汉族人的GSTM1纯合缺失频率为56.1%,GSTP1105I/I、I/V和V/V基因型频率分别为60.7%、35.2%和4.1%。杂合I/V基因型组的平均GST酶活性(3.53±0.63U·g-1Hb)比野生I/I基因型的GST酶活性低(4.25±1.07U·g-1Hb,P=0.000),比突变V/V基因型的GST酶活性高(2.44±0.67U·g-1Hb,P=0.004)。在GSTM1(-)基因组,GSTM1(-)/GSTP1I/I基因型携带者的GST酶活性比GSTM1(-)/GSTP1I/V或V/V携带者的高,而在GSTM1(+)组,两组间的酶活性无差异。不同年龄组平均GSTs活性无差异,而女性的平均GSTs活性比男性的平均值高,但差异无显著性。结论尽管其它GST酶可能会稀释GSTP1基因型对GST酶活性的效果,GST酶活性仍与GSTP1105Val基因型呈很强的相关性。

CpG寡核苷酸的筛选及其对rSj28GST的免疫佐剂作用

目的 研究 Cp G寡核苷酸 (Cp G ODN)对日本血吸虫重组 2 8k Da GST(r Sj2 8GST)抗原分子的免疫佐剂作用和诱导 Th1型反应的效应。方法 用淋巴细胞增殖试验筛选对鼠有良好免疫刺激作用的 Cp G ODN;用基因工程方法生产 r Sj2 8GST抗原分子 ;EL ISA检测小鼠抗 r Sj2 8GST的同型抗体 Ig G1 、Ig G2 a水平。结果 筛选出的 Cp G ODN如 182 6、2 0 0 2、182 6 4、2 10 2等具有良好的免疫刺激活性 ,其 SI>2 ;纯化的 r Sj2 8GST抗原分子 ,呈单一电泳条带 ;以 Cp G ODN为 r Sj2 8GST免疫佐剂 ,其免疫小鼠产生的抗体滴度显著高于对照组 ,且同型抗体 Ig G2 a与 Ig G1 比值也明显升高。结论 Cp G ODN182 6 4是一种良好的免疫佐剂 ,并可显著诱导

响应面设计法优化GST发酵培养基

利用响应面方法对重组谷胱甘肽硫转移酶表达菌株E coliBL21(DE3)PGEX发酵生产谷胱甘肽硫转移酶(GST)的培养基进行了优化。用Plackett-Burman实验方法研究葡萄糖、酵母膏和MgSO4等20个营养因子对产GST活力的影响,结果表明主要影响因子为葡萄糖、酵母膏和MgSO4。根据实验结果对主要影响因子的浓度范围进行估计,然后用Box-Behnken设计及响应面分析确定主要影响因子的最佳浓度。结果表明当葡萄糖浓度为42.06 g/L,蛋白胨浓度为10g/L,酵母膏浓度为8.47 g/L,NaCl浓度为1 g/L,MgSO4浓度为1.62 g/L时,E coliBL21(DE3)PGEX产GST活力达到633.9 mmol/(L.h),较原始培养基的活力提高了63.75%。

GSTM1、GSTT1基因多态性和饮酒习惯与原发性肝癌发生的危险性

目的 研究GSTM 1和GSTT 1基因的多态性和饮酒习惯与原发性肝癌发生的危险性。方法 在肝癌高发区江苏省泰兴市进行了以人群为基础的病例对照研究 ,并应用多重PCR法检测了 2 0 7例原发性肝癌及其 1∶1配对的正常对照的GSTM 1和GSTT 1基因型。结果 GSTM 1空白基因型的频率 ,病例组为 5 8.94% ,对照组为 5 7.0 0 % ;GSTT 1空白基因型的频率 ,病例组和对照组分别为 5 2 .17%和 46.86% ,2组间无显著性差异。但GSTT 1的空白基因型与非空白基因型相比 ,当其长期饮用高度白酒达2 3年以上或月饮酒量大于 3 0 0 0 g时 ,患肝癌的危险性显著增高 (OR =2 .5 6,95 %CI为 1.0 8～ 6.0 5或OR =3 .48,95 %CI为 3 1.47～ 8.2 2 ) ;此外分析饮酒总量 (kg·年 )也得到同样的结果 (OR =3 .71,95 %为 1.5 1～ 9.12 )。结论 携带GSTT 1空白基因型且有长期大量饮酒习惯者 。

木豆抗旱相关基因CcGST1克隆与表达分析

为探讨谷胱甘肽转移酶(GST)的抗逆机理,克隆木豆中的抗旱谷胱甘肽转移酶基因,基于木豆干旱胁迫c DNA文库中1个GST基因相关的EST片段,利用RACE技术从木豆幼苗中克隆了该基因,命名为CcGST1,研究其在不同组织及不同干旱时期的表达特性。结果表明:该基因含1个669 bp的开放阅读框,编码223个氨基酸残基,含有保守的谷胱甘肽-S-转移酶的N端和C端结构域;系统进化树分析结果表明木豆CcGST1与大豆、宽叶菜豆等豆科植物的GST亲缘关系较近;亚细胞定位预测CcGST1蛋白可能定位于叶绿体;荧光定量PCR分析表明,CcGST1在木豆幼苗根、茎和叶中均有表达,但根中的表达水平最高,同时该基因受干旱胁迫诱导表达。据此推测木豆CcGST1基因可能与木豆应激干旱胁迫相关。

二斑叶螨抗螺螨酯品系GST基因的克隆与表达分析

【目的】揭示二斑叶螨Tetranychus urticae对螺螨酯的分子抗性机理。【方法】利用RT-PCR克隆了二斑叶螨的谷胱甘肽-S-转移酶(glutathione S-transferase,GST)基因cDNA全长序列,采用生物信息学软件分析了克隆基因的编码蛋白特性;利用实时荧光定量PCR方法分析GST基因在二斑叶螨的螺螨酯抗性与敏感品系中的表达差异。【结果】克隆获得的谷胱甘肽-S-转移酶2个基因分别被命名为TuGSTd1和TuGSTd2(GenBank登录号分别为:KC445659和KC445660)。序列分析发现,TuGSTd1的开放阅读框长度为648bp,编码215个氨基酸,分子量约为24.47kDa,理论等电点为5.49;TuGSTd2的开放阅读框为648bp,编码215个氨基酸,分子量约为24.57kDa,理论等电点为6.33。系统发育分析表明这两个基因与桔全爪螨Panonychus citri Delta家族的GST基因的氨基酸序列一致性为93%。实时荧光定量PCR结果表明,TuGSTd1和TuGSTd2在二斑叶螨抗螺螨酯品系中的相对表达量分别为敏感品系的5.60和3.75倍。【结论】GST基因在二斑叶螨抗螺螨酯品系中的相对表达量均显著高于敏感品系,据此推测GST基因的过量表达可能与其对螺螨酯的抗性形成有关。