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樟疫霉多聚半乳糖醛酸酶基因9和10的克隆、测序及其遗传转化研究 被引量：6

Studies on cloning,sequencing and genetic transformation of Pcpg (Phytophthora cinnamomi polygalac- turonase) 9 and Pcpg 10
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摘要 　研究优化了樟疫霉多聚半乳糖醛酸酶基因9(Pcpg9)和Pcpg10基因克隆的PCR条件,并对其进行了克隆、测序和遗传转化研究。克隆获得了2个基因,其大小前者1059bp,后者1290bp;通过构建表达载体和遗传转化获得了2个基因的转基因菌系;2个基因均能指导合成相应的PG酶蛋白,其中转化Pcpg10基因的酵母菌所分泌的PG酶活性最强,对照Anpg 的PG酶活性次之,Pcpg9的PG酶活性最弱。Westernblotting表明,所克隆的2个基因所指导合成的PG酶蛋白均有不同程度的糖基化。 Pcpg (Phytophthora cinnamomi polygalacturonase) 9 and Pcpg 10 genes were cloned based on the studying of the PCR composition and its reaction condition of the genes.The sequence and their expression vectors of the genes were done and the genes were transformed to a yeast line of W303-1B.Then the PG activity of transgenic lines was investigated.The genes were cloned and the size of Pcpg 9 was ( 1 059 bp) and Pcpg 10 was 1 290 bp.The transgenic yeast lines of Pcpg 9 and Pcpg 10 genes were obtained respectively.Pcpg 9,Pcpg 10 and the control,Anpg(Aspergillus niger polygalacturonase) Ⅰ could guide to synthesize the PGs.The PG activity of transgenic line of Pcpg 10 was the strongest,and that of the Pcpg 9 gene was the weakest.Western blotting results showed that there were different degree glycosylation of PGs which were encoded by Pcpg 9 and Pcpg 10 genes.
作者 巩振辉 Arvid Gotesson David A Jones
机构地区 西北农林科技大学园艺学院 Plant Cell Biology
出处 《西北农林科技大学学报（自然科学版）》 CSCD 北大核心 2004年第8期1-6,共6页 Journal of Northwest A&F University(Natural Science Edition)
基金 中澳合作项目 陕西省自然科学基金项目(2001SM11) 陕西省农业分子生物学重点实验室基金项目
关键词 疫病病原菌 Pcpg基因 克隆 测序 遗传转化 PG活性 Phytophthora cinnamomi Pcpg gene cloning sequencing genetic transformation PG activity
分类号 Q785 [生物学—分子生物学]
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参考文献22

  • 1De veaal E J I,Grous K C,Huber D J,et al.Degradation and solubilization of pectin by -galactosidases purified from arocado mesocarp[J].Physiol Plant,1993,87:279-285.
  • 2Cole A L J.Pectin enzyme activity from Phytophthora infestans[J].Phytochemistry,1969,9:337-340.
  • 3Isshiki A,Akimitsu K,Nishio K,et al.Purfication and characterization of an endopolygalacturonase from the rough lemon pathotype of Alternaria alternata,the cause of citrus brown spot disease[J].Physiological and Molecular Plant Pathogy,1997,52:155-167.
  • 4Jarvis M C,Threlfall D R,Friend J.Potato cell wall polysaccharides:Degradation with enzymes from Phytophthora infestans[J].Journal of Experiment Botany,1981,32:1309-1319.
  • 5Lei S P,Lin H C,Heffernan L,et al.Evidence that polygalacturonase is a virulence determinant in Erwinia carotovora[J].Journal of Bacteriology,1985,164:831-835.
  • 6Martel M B,Letoublon R,Fevre M.Purification and characterization of two endopoly- galacturonases secreted during the early stages of the saprophytic growth of Sclerotinia sclerotiorum[J].FEMS Microbiology Letter,1998,158:133-138.
  • 7Martel M B,Letoublon R,Fevre M.Purification of endopolygalacturonases from Sclerotinia sclerotiorum:Multiplicity of the complex enzyme system[J].Current Microbiology,1996,33:243-248.
  • 8Panabieres F,Marais A,Le Berre J Y,et al.Characterization of a gene cluster of Phytophthora cryptogea which codes for elicitins,proteins inducing a hypersensitive-like response in tobacco[J].Mo1 Plant-Microbe Interact,1995,8:996-1003.
  • 9Pieterse C M J,Van West P,Verbakel H M,et al.Structure and genomic organization of the ipiB and ipiO gene clusters of Phytophthora infestans[J].Gene,1994,138:67-77.
  • 10Van Santen Y,Benen J A E,Schroter K H,et al.1.68-(A) crystal structure of endopolygalacturonase Ⅰ from Aspergillus niger and identification of active site residues by site-directed mutagenesis[J].J Biol Chem,1999,274:30474-30480.

同被引文献91

  • 1巩振辉,吕元红,王晓敏.樟疫霉菌多聚半乳糖醛酸酶16和17基因的克隆、测序及其真核表达研究[J].西北农林科技大学学报（自然科学版）,2005,33(6):1-6. 被引量：9
  • 2沈崇尧 等.中国大豆疫霉根腐病的发现及初步研究[J].植物病理学报,1991,21(4):298-298.
  • 3SCHMITHENNER A F. Problems and progressing in control of Phytophthora root rot of soybean [J]. Plant Disease,1985,69:462-468.
  • 4MAR.TEL M B,LETOUBLON R,FEVRE M. Purification and characterization of two endopolygalacturonases secreted during the early stages of the saprophytic growth of Sclerotinia sclerotiorum [ J ]. FEMS Lett, 1998, 158: 133 - 138.
  • 5SAKAMOTO T,BONNIN E,QUEMENER B,et al. Purification and characterisation of two exo-polygalacturonases from Aspergillus niger able to degrade xylogalacturonan and acetylated homogalacturonan[J]. Biochimica et Biophysica Acta,2O02,1572( 1 ) : 10 - 18.
  • 6SALCH A,NEVIN M F,EBTSANA N H,et al. Biochemical characterization of an extracellular polygalacturonase form Trichderma harzianum[J]. Journal of Biotechnology ,2006 ,127 :54 -64.
  • 7SHIEH M I,BROWN R L,WHITEHEAD M P,et at. Molecular genetic evidence for the involvement of a specific polygalacturonase,Pc2,in the invasion and spread of Aspergillus flavus in cotton bolls[J]. Appl Environ Microbiol, 1997,63:3548 - 3552.
  • 8ISSHIKI A,AKIMITSU K,YAMAMOTO M,et al. Endopolygalacturonase is essential for citrus black rot caused by Alternaria cirri but not brown spot caused by Alternaria alternata [ J ]. Mol Plant-Microbe Interact, 21301,14 : 749 - 761.
  • 9LI J, GOODWIN P H, Expression of cgmpg2,an endopolygalacturonase gene of Colletotrichum gloeosporioides f. sp. Malvae, in culture and during infection of Malva pusilla[ J ]. J Phytopathology ,2002,150:213 -219.
  • 10COTTON P,RASCLE C ,FEVRE M,et al. Characterization of PG2,an early endo PG produced by Sclerotinia sclerotiorum, expressed in yeast [ J ]. FEMS Microbiology Letters ,2002,213:239 - 244.

引证文献6

二级引证文献20

西北农林科技大学学报（自然科学版）
2004年 第8期

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