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杆状病毒AcMNPV p35基因的克隆表达及多抗制备 被引量：1

Cloning and Expression of Autographa californica Nucleopolyhedrovirus p35 Gene in E. coli and Preparation of Multiclonal Antibody
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摘要 用PCR方法扩增得到苜蓿丫纹夜蛾核多角体病毒(Autographa californic anucleopolyhedrovirus,AcM N-PV)p35基因,将其克隆至质粒pET-32a(+)上,构建得到重组质粒pET-p35,转化大肠杆菌BL21(DE3),经IPTG诱导,表达了1条约为55 ku的蛋白带。以Ni2+-NTA偶连抗体检测证明所表达的蛋白为带有组氨酸的融合蛋白。采用割胶回收的方法纯化融合蛋白,以纯化的融合蛋白制备多克隆抗体,效价为1/1 024。免疫印迹分析表明,该抗血清能与感染苜蓿丫纹夜蛾核多角体病毒的细胞蛋白样品发生特异性反应。 Autographa californica nucleopolyhedrovirus （AcMNPV） p35 gene was amplified from AcMNPV genomic DNA via PCR. The PCR product was cloned into the expression vector pET-32a（ ＋ ） and transformed into Escherichia coli BL21 （DE3） , and a protein around 55 ku was expressed after induction with IPTG （ isopropylthio-β-D-galaetoside）. Western blot using Ni-NTA Conjugate indicated that the protein contained 6 x His tag. The fusion protein was separated on SDS-PAGE and recovered by gel extraction, and used as the immunogen to raise multielonal antibody, which had a titer of 1/1 024. Western blotting analysis indicated that the antibody could react with the corresponding protein existed in AeMNPV-infeeted cells.
作者 李玲玲 李朝飞 庞义
机构地区 华南师范大学激光生命科学教育部重点实验室 中山大学生物防治国家重点实验室
出处 《微生物学杂志》 CAS CSCD 2007年第5期1-4,共4页 Journal of Microbiology
基金 国家重大基础研究项目(973项目 G2000016209)
关键词 苜蓿丫纹夜蛾核多角体病毒p35基因基因表达多克隆抗体 Autographa californica nueleopolyhedrovirus p35 gene prokaryotie expression multielonal antibody
分类号 Q93 [生物学—微生物学]
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参考文献15

  • 1Krakaue D C, Payne R J. The evolution of virus-induced apoptosis[J]. Proc Biol Sci, 1997, 264(1 389) :1 757-1 762.
  • 2Clem R .I, Fechheimer M, Miller L K. Prevention of apoptosis by a baculovirus gene during infection of insect cells[J]. Science, 1991, 254(5 036) :1 388-1 390.
  • 3南方,宋建华,王瑞青,梁昌镛,陈新文.AcMNPV P35抑制HaSNPV诱导的Tn-Hi5细胞凋亡[J].中国病毒学,2005,20(2):173-178. 被引量：2
  • 4Sugimoto A, Friesen P D, Rathran J H, et al. Baculovirus p35 prevents developmentally programmed cell death and rescues a ced-9 mutant in the nematode Caenorhabditis elegans[J]. EMBO J, 1994, 13(9) :2 023-2 028.
  • 5Hay B A, Wolff J, Rulin G M, et al. Expression of baculovirus p35 prevents cell death in Drosophila [ J]. Development, 1994, 120(8):2 121-2 129.
  • 6Rabizadeh S, LaCount D J, Friesen P D, et al. Expression of the baculovirus p35 gene inhibits mammalian neural cell death [J]. J Neurochem, 1993, 61(6) :2 318-2 321.
  • 7Sah NK, Taneja TK, Pathak N, et al. The baculovirus anti-apoptotic p35 gene also functions via an oxidant-dependent pathway[J]. PNAS, 1999, 96(9) :4 838-4 843.
  • 8Priya R, Punya S, Sukh M S, et al. Baculovirus p35 inhibits NO-induced apoptosis in activated macrophages by inhibiting cytochrome c release[J].J Cell Sci, 2004, 117(14) :3 031- 3 039.
  • 9曾季平,王立祥,胡晓燕,秦文,孔峰,杨玲玲,崔行,刘新垣.p35基因对MnCl_2诱导PC12细胞凋亡的作用[J].山东大学学报（医学版）,2004,42(6):625-628. 被引量：6
  • 10Hershberger P A, LaCount D J, Friesen P D. The apoptotic suppressor p35 is required early during baculovirus replication and is targeted to the cytosol of infected cells [ J]. J Virol, 1994, 68(6) :3 467-3 477.

二级参考文献28

  • 1Vilaplana L, O'Reilly D R. Functional interaction between Cydia pomonella granulovirus IAP proteins [J]. Virus Res, 2003,92:107-111.
  • 2Blissard G W. Baculovirus-insect cell interactions [J]. Cytotechnology, 1996,20:73-93.
  • 3Gozani O, Boyce M, Yoo L, et al. Life and death in paradise [J]. Nat Cell Biol ,2002,4:E159-162.
  • 4Krakaue D C, Payne R J.The evolution of virus-induced apoptosis [J]. Proc R Soc Lond B Biol Sci, 1997,264:1757-1762.
  • 5Kamita S G, Majima K, and Maeda S. Identification and characterization of the p35 gene of Bombyx mori nuclear polyhedrosis virus that prevents virus-induced apoptosis [J]. J Virol, 1993,67:455-463.
  • 6Ishikawa H, Ikeda M, Yanagimoto K, et al. Lavina-Caoili, and M. Kobayashi.Induction of apoptosis in an insect cell line, IPLB-Ld652Y, infected with nucleopolyhedroviruses[J]. J Gen Virol, 2003,, 84:705-714.
  • 7Clarke T E, and Clem R J. In vivo induction of apoptosis correlating with reduced infectivity during baculovirus infection [J]. J Virol, 2003, 77:2227-2232.
  • 8Clem R J, Fechheimer M,Miller L K. Prevention of apoptosis by a baculovirus gene during infection of insect cells [J]. Science, 1991, 254:1388-1390.
  • 9del Pozo O,Lam E. Expression of the baculovirus p35 protein in tobacco affects cell death progression and compromises N gene-mediated disease resistance response to Tobacco mosaic virus [J]. Mol Plant Microbe Interact, 2003,16:485-494.
  • 10Sugimoto A, Friesen P D,Rothman J H. Baculovirus p35 prevents developmentally programmed cell death and rescues a ced-9 mutant in the nematode Caenorhabditis elegans[J]. Embo J, 1994, 13:2023-2028.

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微生物学杂志
2007年 第5期

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