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龙眼GPX基因的克隆、原核表达及其在体胚发生过程中的表达分析 预览 被引量:2

Cloning of GPX, Prokaryotic Expression and Its Expression During Longan Somatic Embryogenesis of Dimocarpus longan Lour.
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摘要 采用RT-PCR与RACE相结合的方法.从龙眼胚性愈伤组织中克隆了长947bD含有完整开放阅读框的龙眼D1GPXcDNA序列(GenBank登录号为EU364813)和长度为1736bp的DNA序列(GenBank登录号为EU680970)。其编码1个含有168个氨基酸的蛋白质。DIGPX基因中含有5个内含子,均符合真核生物内含子通用的GT—AG法则。生物信息学分析显示:该蛋白为亲水的酸性蛋白质,定位于叶绿体:与其他植物的GPX有较高的同源性。将该基因构建成原核表达载体.经IirfG诱导表达了1个分子量约为23ku的蛋白。利用实时荧光定量PCR(qPCR)技术研究该基因在龙眼体胚发生发育过程中的表达情况.结果显示.从松散型胚性愈伤组织到不完全胚性紧实球形结构阶段,D1GPXmRNA的转录水平逐渐升高,在胚性紧实球形结构阶段降到最低水平.子叶形胚阶段又上升到与不完全胚性紧实球形结构阶段相当的水平。 Using RT-PCR and RACE, DIGPX, a 947 bp cDNA with an ORF was isolated from the embryogenic calli of Dimocarpus longan. The DlGPX gene, encoding a protein with 168 amino acids, contained 5 introns consistent with the GT-AG dogma for eukaryotic introns. Bioinformatic analysis indicated that DlGPX was a hydrophilic acidic protein and located in the chloroplast. Its amino acid sequence showed high similarity with D1GPX from other plant~ Prokaryotic expression vector fused with DIGPX was constructed and a protein about 23 ku was expressed. qPCR results indicated that the transcription level of DlGPX increased gradually from embryogenic calli to incomplete compact pro-embryogenic culture stage and decreased to the lowest level at compact pro-embryogenic culture stage, then upregulated to a comparable level with that at incomplete compact pro-embryogenic culture stage.
作者 陈义挺 赖钟雄 方智振 蔡英卿 林玉玲 李焕苓 陈登云 CHEN Yiting1'2, LAI Zhongxiong1, FANG Zhizhen1, CAI YingqingI LIN Yulingl, LI Huanlingt, CHEN DengyunI 1 Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China 2 Institute of Fruit, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China
出处 《热带作物学报》 CSCD 北大核心 2013年第8期1483-1489,共7页 Chinese Journal of Tropical Crops
基金 国家自然科学基金项目(No.31071787、31201588、30471204) 高等学校博士学科点专项科研基金项目(N0.20093515110005):福建省科技平台建设项目(No.2008N2001).
关键词 龙眼 体细胞胚胎发生 GPX 克隆 生物信息学 实时荧光定量PCR Longan Somatic embryogenesis GPX Cloning Bioinformatics qPCR
作者简介 陈义挺(1972年-),男,博士,副研究员;研究方向:果树生物技术与遗传资源。 通讯作者:赖钟雄,E-mail:laizx01@163.com。
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  • 1Gaber A,Ogata T,Maruta T,et al.The Involvement ofArabidopsis Glutathione Peroxidase 8 in the Suppression ofOxidative Damage in the Nucleus and Cytosol [J].Plant AndCell Physiology,2012,53(9):1 596-1 606. 被引量:1
  • 2Margis R,Dunand C,Teixeira F K,et al.Glutathione peroxidasefamily-an evolutionary overview [J].FEBS Journal,2008,275(15):3 959-3 970. 被引量:1
  • 3刘家忠,龚明.植物抗氧化系统研究进展[J].云南师范大学学报:自然科学版,1999,19(6):1-11. 被引量:61
  • 4Maiorino F M,Brigelius-Floh6 R,Aumann K,et al.Diversityof glutathione peroxidases [J].Methods in Enzymology,1995,252:38-53. 被引量:1
  • 5Arthur J.The glutathione peroxidases[J].Cellular And MolecularLife Sciences,2000,57(13):1 825-1 835. 被引量:1
  • 6Jung B G,Lee K 0,Lee S S,et al.A Chinese cabbagecDNA with high sequence identity to phospholipid hydroperoxideglutathione peroxidases encodes a novel isoform of thioredoxin-dependent peroxidase[J].Journal of Biological Chemistry,2002,277(15):12 572-12 578. 被引量:1
  • 7Criqui M,Jamet E,Parmentier Y,efaL Isolation and characterizationof a plant cDNA showing homology to animal glutathioneperoxidases[J].Plant Molecular Biology,1992,18(3):623-627. 被引量:1
  • 8Eshdat Y,Holland D,Faltin Z,et al.Plant glutathioneperoxidases[J].Physiologia Plantarum,1997,100(2):234-240. 被引量:1
  • 9Sugimoto M,Furui S,Suzuki Y.Molecular cloning andcharacterization of a cDNA encoding putative phospholipidhydroperoxide glutathione peroxidase from spinach[J].BiosciencesBiotechnology and Biochemistry,1997,61(8):1 379-1 381. 被引量:1
  • 10Sugimoto M,Sakamoto W.Putative phospholipid hydroperoxideglutathione peroxidase gene from Arabidopsis thaliana inducedby oxidative stress[J].Genes & Genetic Systems,1997,72(5):311-316. 被引量:1

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同被引文献43

  • 1苗雨晨,白玲,苗琛,陈珈,宋纯鹏.植物谷胱甘肽过氧化物酶研究进展[J].植物学通报,2005,22(3):350-356. 被引量:41
  • 2Ausubel F.M., Kingston R.E., Brent R., Seidman J.G., Moore D.D., Smith J.A., and Struhl K., eds., Yan Z.Y., and Wang H.L., trans., 1999, Short Protocols in Molecular Biology, Science Press,Beijing,China,pp.700(F.M.奥斯伯,R.E.金斯顿,R.布伦特,J.G.塞德曼,D.D.穆尔,J.A.史密斯,K.斯特拉尔,主编,颜子颖,王海林,主译,1999,精编分子生物学实验指南,科学出版社,中国,北京,pp.700) 被引量:2
  • 3Smimoff N. Environment and plant metabolism. Oxford: Bins Scientific Publishers, 1995:217-243. 被引量:1
  • 4Sugimoto M, Sakamoto W. Putative phospholipid hydroperoxide glutathione peroxidase gene from Arabidopsis thaliana induced by oxidative stress. Genes & Genetic Systems, 1997, 72(5): 311-316. 被引量:1
  • 5Criqui M C, Jamet E, parmentier Y, Marbach J, Durr A, Fleck J. Isolation and characterization of a plant eDNA showing homology to animal glutathione peroxidases. Plant Molecular Biology, 1992, 18: 623-627. 被引量:1
  • 6Li W J, Feng H, Fan J H, Zhang R Q, Zhao N M, Liu J Y. Molecular cloning and expression of a phospholipid hydroperoxide glutathione peroxidase homolog in Oryza sativa. Biochim Biophys Acta, 2000, 1493(1/2): 225-230. 被引量:1
  • 7Depege N, Drevet J, Boy~r N. Molecular cloning and characterization of tomato cDNAs encoding glutathione peroxidase-like proteins. European Journal of Biochemistry, 1998, 253(2): 445-451. 被引量:1
  • 8Sugimoto M, Furui S, Suzuki Y. Molecular cloning and characterization of a eDNA encoding putative phospholipid hydroperoxide glutathione peroxidase from spinach. Biosciences Biotechnology and Biochemistry, 1997, 61(8): 1379-1381. 被引量:1
  • 9Avsian-Kretehrner O, Eshdat Y, Gueta-Dahan Y, Ben-Hayyim G. Regulation of stress-induced phospholipid hydroperoxide glutathione peroxidase expression in citrus. Planta, 1999, 209(4): 469-477. 被引量:1
  • 10陈义挺,王卫,陈婷.福建省地方梨种质资源莆田豆梨GPX基因克隆与生物信息学分析.福建省农业科学院青年科技人才创新基金学术论文集.2011:200.207. 被引量:1

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