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山羊痘病毒P32基因真核表达载体的构建及表达 被引量：1

Construction and Expression of Recombinant Antigen P32 Gene of Goatpox Virus
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摘要 从重组克隆载体pMD18-P32中扩增出山羊痘病毒P32基因,与毕赤酵母分泌性表达载体pPIC9K相连接,构建重组表达载体pPIC9K-P32。重组质粒pPIC9K-P32用SalⅠ线性化后,与毕赤酵母(Pichia pastoris)GS115混合后电转化,使重组表达载体与酵母染色体发生同源重组。采用G418抗性梯度筛选法得到高拷贝重组菌株,甲醇诱导目的基因表达。SDS-PAGE和Western-blotting分析结果表明,用酵母成功表达出了31 kD的重组蛋白,该蛋白具有生物学活性,能被山羊痘阳性血清识别。 The structural protein P32 gene of the goat pox virus in the recombinant plasmid pMD18-P32 was subcloned into the Pichia pastoris expression vector pPIC9K. The resultant recombinant plasmid pPIC9K-P32 was transformed into P. pastoris GSl15 by electroporation. The multi-copy recombinant P. pastoris strains were screened by G418 and induced by methanol. The expressed product was analyzed by SDS-PAGE and Western-blotting tests. The results indicated that the recombinant protein was expressed in P. pastoris, and had biological activity,the protein could be recognized by anti-serum against goat pox virus.
作者 谢芳 张强 颜新敏 吴国华 李健 朱海霞 郭爱疆 韩若婵 施程洪 鞠厚斌 朱彩珠 岳城 才学鹏
机构地区 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部畜禽病毒学重点开放实验室 新疆农业大学动物医学学院
出处 《新疆农业大学学报》 CAS 2008年第3期1-4,共4页 Journal of Xinjiang Agricultural University
基金 国家基础平台建设项目(2005DKA21205-3)
关键词 山羊痘病毒 P32基因 毕赤酵母 诱导表达 Goatpoxvirus P32 gene pichia pastoris induction expression
分类号 S851.654 [农业科学—预防兽医学]
  • 相关文献

参考文献16

  • 1Cran V M. Control of capripoxvirus infection[J]. Vaccine, 1993,11 (3) : 1275-1279.
  • 2方明祥,苏飞,王玉瑾,高以武,朱涛,胡长文.山羊痘的诊治[J].畜牧与兽医,2003,35(3):45-45. 被引量：38
  • 3Paul D Gershon. A comprison of rhe Genome Organization of Capripoxvirus with That of the Orthopoxviruses[J]. Journal of Viroogl, 1989,11:4703-4708.
  • 4Distt Nainital. A comprehensive reviews of goat pox and sheep pox and their diagnosis[J]. Animal Health Research Reviews,2000,1 (2) : 127-136.
  • 5V M Carn. Protection of goats against capripox using a subnit vaccine[J]. The Veterinary Record, 1994,26:434-436.
  • 6E R Tulman. The Genomes of Sheepox and Goatpox Viruses[J]. Virology, 1995,209(1) :207-12.
  • 7R P Kitching. Studies on the Major Common Precipitatation Antigen of Capripoxvirus[J]. Virology, 1986,67:139-148.
  • 8Paul D Gershon. A comparison of the Genomes of Capripoxvirus Isolates of Sheeps, Goat, and CattleEJ~. Virology, 1988, 134:341-9.
  • 9Chand P. Western blot analysis of virus specific antibody for capripox and contagious pustular dermatitis viral infections in sheep[J]. Epidemiology and Infection,1994,113(2):377-385.
  • 10H G Heine, M P Stevens. A capripoxvirus detection PCR and antibody ELISA based on the major P32, the homolog of the vaccinia virus H3L gene[J]. Journal of Immunolofical Methods, 1999,227:187-196.

二级参考文献11

  • 1Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd. New York: Cold Spring Harbor Laboratory Press, 1989. 1.25～1.28
  • 2Gietz R D, Schiestl R H. Transforming yeast with DNA. Meth Mol Cell Biol, 1995, 5(5): 255-269
  • 3Plessis A, Dujon B. Multiple tandem integrations of transforming DNA sequences in yeast chromosomes suggest a mechanism for integrative transformation by homologous recombination. Gene, 1993, 134(1): 41～50
  • 4Adams A, Gottschling D E, Kaiser C A, et al. Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual. New York: Cold Spring Harbor Laboratory Press, 1998. 35～36
  • 5Kruckeberg A L, Ye L, Berden J A, et al. Functional expression, quantitation and cellular localization of the Hxt2 hexose transporter of Saccharomyces cerevisiae tagged with the green fluorescent protein. Biochem J, 1999, 339(Pt 2): 299～307
  • 6Baere T D, Claeys G, Swinne D, et al. Identification of cultured isolates of clinically important yeast species using fluorescent fragment length analysis of the amplified internally transcribed rRNA spacer 2 region. BMC Microbiol, 2002, 2(1): 21～27
  • 7Yamada Y,Makimura K, Merhendi H,et al. Comparison of different methods for extraction of mitochondrial DNA from human pathogenic yeasts. Jpn J Infect Dis, 2002, 55(4): 122～125
  • 8Millar B C, Jiru X, Moore J E,et al. A simple and sensitive method to extract bacterial, yeast and fungal DNA from blood culture material. J Microbiol Methods, 2000, 42(2):139～147
  • 9Haaning J, Oxvig C, Overgaard M T, et al. Simple and reliable procedure for PCR amplification of genomic DNA from yeast cells using short sequencing primers. Biochem Mol Biol Int, 1997, 42(1):169～172
  • 10Deak T, Chen J, Beuchat L R. Molecular characterization of Yarrowia lipolytica and Candida zeylanoides isolated from poultry. Appl Environ Microbiol, 2000, 66(10): 4340～4344

共引文献71

同被引文献25

  • 1康文玉,徐自忠,高洪,花群义,周晓黎,杨云庆,董俊.羊痘病毒[J].中国畜牧兽医,2004,31(12):33-36. 被引量：18
  • 2郭洪勋,张加勇,赵晓春,孙刚,李鑫,王宇祥.PCR检测山羊痘病毒试验的建立[J].畜牧兽医科技信息,2005,21(4):42-42. 被引量：4
  • 3许乐仁,周碧君,阴正兴,何光志,刘忠伟.山羊痘病变的超微病理学研究[J].畜牧兽医学报,2005,36(5):492-497. 被引量：28
  • 4马维民,刘湘涛,张强,吴国华,颜新敏,李健,朱海霞,吴润.山羊痘病毒P32基因的克隆与序列分析[J].甘肃农业大学学报,2006,41(4):27-30. 被引量：19
  • 5Alcami A, Keszinowski U H. Viral mechanisms of immune era' s ion[J]. Trends Microbiol, 2000,8 : 410 - 418.
  • 6Carn V M,Timms C P,et al. Protection of goats against capripox using a subunit vaccine[J]. Veterinary Record, 1994,135( 18): 434-436.
  • 7Carn V M. An antigen trapping ELISA for the detection of capripoxvirus intisse culture supernatant biopsy samples[J]. Journal of Virological Methods, 1995,49:285 - 294.
  • 8Chand P. Western blot analysis of virus specific antibody for capripox and contagious pustular dermatitis viral infections in sheep [J]. Epidemiology and Infection,1994,113(2):377-385.
  • 9Gershon P D,Black D N. A comparison of the genomes of Capripoxvirus isolates of sheep,goats,and cattle [J]. Virology, 1988, 164:341-349.
  • 10Heine H G,Stevens M P,Foord A J,et al. A capripoxvirus detection PCR and antibody ELISA based on the maior antigen P32, the homolog of the vaccinia virus H3L gene [J]. J Immune Meth,1999,277: 187-196.

引证文献1

二级引证文献16

新疆农业大学学报
2008年 第3期

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