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狂犬病病毒SRV_9株修饰全长基因组cDNA克隆的构建 被引量:2

Construction of Full-Length Genomes-Modified cDNA Clone for Rabies Virus Strain SRV_9
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摘要 为获得狂犬病病毒(rabies virus,RV)SRV9株的基因组全长cDNA克隆并对其进行基因修饰,深入研究狂犬病病毒的基因组特性及构建新型病毒载体,本研究根据GenBank中公布的SRV9基因组序列设计数对引物,以SRV9病毒为材料,从病毒总RNA中将全长基因组11 928 bp分5段扩增,克隆至载体测序鉴定。测序正确后,设计引物缺失基因组Ψ区并在缺失位置插入人细胞色素C基因,又设计引物删除糖蛋白(G)胞质区(CD区)。利用引物在病毒基因组起始处添加锤头状核酶序列,在病毒序列结尾处添加了丁型肝炎核酶序列。再次测序后克隆至pcDNA3.1(+)载体,利用扩增片段自身内切酶位点顺次进行修饰后全长连接,从而获得含修饰SRV9全长基因组的质粒pcDNA3.1-SRV9,为RV感染性克隆的建立奠定了基础。 In order to obtain full-length genomes-modified cDNA clone for rabies virus strain SRV9 and to deeply study genomic characteriitics of SRV9 and construct the new virus vector and several pairs of primers were designed according to the full-length genomic sequence of SRV9 published on GenBank by using the total RNA extracted from rabies virus as a template.Five DNA fragments covering the entire genome 11 928 bp were amplified,and inserted into vector and were determined for sequeneing.After identification of the sequence,Ψ region of genome was designed with a lack of primers before the human cytochrome c gene was cloned into such a region,and cytoplasmic region of glycoprotein gene was also deleted by primers designed ahead.Hammerhead ribozyme sequence was inserted into the head region of full-length genomes and hepatitis delta virus ribozyme sequence was added into the end region of full-length genomes with the help of primers.Fragments were all cloned into pcDNA3.1(+) vector after sequenced correctly.These five fragments were ligated by using amplified fragments that contain their own restriction enzyme sites to obtain the full-length genomes-modified plasmid clone pcDNA3.1-SRV9,which would contribute to the foundation of infectious clone for rabies virus.
出处 《新疆农业大学学报》 CAS 北大核心 2010年第5期416-421,共6页 Journal of Xinjiang Agricultural University
基金 新疆维吾尔自治区高技术研究发展计划项目(200611107)
关键词 狂犬病病毒SRV9株 修饰基因组 全长基因质粒构建 rabies virus strain SRV9 modification genomes construction of full-length genome plasmid
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参考文献17

  • 1Junji Hosokawa-Muto,Naoto Ito,Kentaro Yamada,et al.Characterization of recombinant rabies virus carrying double glycoprotein genes[J].Microbiol.Immunol.,2006,50(3):187-196.
  • 2王朝君.我国狂犬病发病数全球第二[J].中国卫生,2007(12):60-60. 被引量:2
  • 3袁慧君,王三虎,秦鄂德.狂犬病病毒分子生物学研究进展[J].生物技术通讯,2004,15(6):596-599. 被引量:16
  • 4McGettigan J P,Pomerantz R J,Slier C A,et al.Second-generation rabies virus-based vaccine vector expressing human immunodeficiency virus type I gag have greatly reduced pathogenicity but are highly immunogenic[J].J.Viro.,2003,77(1):237-244.
  • 5Schnell M J,Mebatsion T,Corelmann K K.Infectious rabies viruses from Cloned cDNA[J].EMBO,1994,13(18):4195-4203.
  • 6王铁东,张守峰,扈荣良,郭建顺.狂犬病病毒口服疫苗株SRV_9基因组全长cDNA的克隆及特性分析[J].中国兽医学报,2005,25(4):368-370. 被引量:9
  • 7Inoue K I,Shoji Y,Kurane I,et al.An improved method for recovering rabies virus from cloned cDNA[J].Journal of Veterinary methods,2003,107(2):229-236.
  • 8Dietzschold M L,Faber M,Mattis J A,et al.In vitro growth and stability of recombinant rabies viruses designed for vaccination of wildlife[J].Vaccine,2004,23(4):518-524.
  • 9Morimoto K,Foley H D,McGettigan J P,et al.Reinvestigation of the role of the rabies virus glycoprotein in viral pathogenesis using a reverse genetics approach[J].J.Neurovirol.,2000,6(5):373-381.
  • 10魏玉荣,易忠,符子华,马素贞,王晓萍,简子健,魏婕,王海烽,朱晶晶.狂犬病病毒SRV_9株N、P、G和L蛋白基因的克隆及表达载体构建[J].新疆农业科学,2009,46(2):354-358. 被引量:8

二级参考文献67

  • 1王铁东,张守峰,扈荣良,郭建顺.狂犬病病毒口服疫苗株SRV_9基因组全长cDNA的克隆及特性分析[J].中国兽医学报,2005,25(4):368-370. 被引量:9
  • 2ZHENG Haixue,LIU Xiangtao,SHANG Youiun,WU Jinyan,BAI Xingwen,JIN Ye,SUN Shiqi,GUO Huichen,TIAN Hong,FENG Xia,YIN Shuanghui,GUO Jianhong,CONG Guozheng,LIU Zaixin,CHANG Huiyun,MA Junwu,XIE Qingge.Infective viruses produced from full-length complementary DNA of swine vesicular disease viruses HK/70 strain[J].Chinese Science Bulletin,2006,51(17):2072-2078. 被引量:6
  • 3Grubman M J, Baxt B. Foot-and-mouth disease [J]. Clin Microbiol Rev, 2004, 17: 465-93.
  • 4Knipe, David M, Howley, et al. Fields Virology 5th Edition [M]. Lippincott Willians and Wilkins, 2006, 796-839.
  • 5Ryan M D, Belsham G J, King A M. Specificity of enzymesubstrate interactions in foot-and-mouth disease virus polyprotein processing [J]. Virology, 1989, 173: 35-45.
  • 6Brooksby J B, Roger J. Methods of typing and cultivation of foot and mouth disease virus [M]. (Project 208 of OEEC, Paris, 1957): 31-34.
  • 7Valarcher J F, Knowles N J, Fen-is N P, et al. Recent spread of FMD virus serotype Asia 1 [J]. Vet Rec, 2005, 157(1): 30-36.
  • 8Boyer J C, Haenni A L. Infectious transcripts and cDNA clone of RNA virus [J]. Virology, 1994, 198: 415-426.
  • 9Zibert A, Maass G, Strebel K, et al. Infectious foot-and-mouth disease viruses derived from a cloned full-length cDNA [J]. J Virol, 1990, 64: 2467-2473.
  • 10Rieder E, Bunch T, Brown F, et al. Genetically engineered foot- and-mouth disease viruses with poly (C) tracts of two nucleotide are virulent in mice [J]. J Virol, 1993, 67: 5139-5145.

共引文献37

同被引文献13

  • 1袁慧君,王三虎,秦鄂德.狂犬病病毒分子生物学研究进展[J].生物技术通讯,2004,15(6):596-599. 被引量:16
  • 2王铁东,张守峰,扈荣良,郭建顺.狂犬病病毒口服疫苗株SRV_9基因组全长cDNA的克隆及特性分析[J].中国兽医学报,2005,25(4):368-370. 被引量:9
  • 3郭霄峰,富振芳.狂犬病毒糖蛋白基因的重排及病毒的拯救[J].华南农业大学学报,2006,27(1):104-106. 被引量:24
  • 4曾江勇,郭建宏,刘在新.负链RNA病毒的反向遗传技术[J].动物医学进展,2006,27(9):35-39. 被引量:5
  • 5Schnell M J, Mebatsion T,Conzelmann K K. Infectious rabies viruses from cloned cDNA[J]. EMBO, 1994,13 (18) :4195-4203.
  • 6Gao,Y. ,Greenfield, N. J. , Cleverley, D. Z. , & Lenard, J. (1996). The transcriptional form of the phosphoprotein of vesicular stomatitis virus is a trimer: structure and stability. Biochemistry, 35(46) :14,569 -14,573.
  • 7McGettigan, J. P. , Pomerantz, R. J. , Siler, C. A. , McKenna, P. M. , Foley, H. D. , Dietzschold, B. , & Sehnell, M. J. (2003). Sec- ond -generation rabies virus -based vaccine vectors expressing human immunodeficieney virus type 1 gag have greatly reduced pathogenicity but are highly immunogenie. Journal of virology,77( 1 ) :237 -244.
  • 8Morimoto, K. , Hooper, D. C. , Spitsin, S. , Koprowski, H. ,& Dietzschold, B. (1999). Pathogenicity of different rabies virus variants in- versely correlates with apoptosis and rabies virus glycopretein expression in infected primary neuron cultures. Journal of virology, 73( 1 ) :510 -518.
  • 9卢圣栋.现代分子生物学实验技术[M].(第二版)北京:中国协和医科大学出版社,2004.
  • 10杜加亮,唐青.狂犬病病毒反向遗传学及其应用[J].病毒学报,2008,24(6):478-482. 被引量:3

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