期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

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 goat poxvirus
下载PDF
导出
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31000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 poxvirus in the recombinant plasmid pMD18-P32 was subcloned into the Pichia pastoris expression vector pPICgK. The resultant recombinant plasmid pPICgK-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 aim protein was expressed in P. pastoris, and had an immunological activity, the protein could be recognized by anti-serum against goat poxvirus.
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院
出处 《中国兽医学报》 CAS CSCD 北大核心 2009年第8期982-985,共4页 Chinese Journal of Veterinary Science
基金 国家基础平台建设资助项目(2005DKA21205-3)
关键词 山羊痘病毒 P32基因 毕赤酵母 诱导表达 goat poxvirus P32 gene Pichia pastoris induction expression
分类号 S852.65 [农业科学—基础兽医学]
  • 相关文献

参考文献15

  • 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
  • 3Nainital D.A comprehensive reviews of goat pox and sheep pox and their diagnosis[J].Anim Health Res,2000,1 (2):127-136.
  • 4Carn V M.Protection of goats against capripox using a subnit vaccine[J].Vet Rec,1994,26:434-436.
  • 5Tulman E R.The Genomes of sheepox and goatpox viruses[J].Virology,1995,209 (1):207-212.
  • 6Kitching R P.Studies on the major common precipitatation antigen of capripoxvirus[J].Virology,1986,67:139-148.
  • 7Gershon P D.A comparison of the genomes of capripoxvirus isolates of sheeps,goat,and cattle[J].Virology,1988,134:341-349.
  • 8Chand P.Western blot analysis of virus-specific antibody for capripox and contagious pustular dermatitis viral infections in sheep[J].Epidemiol infect,1994,113(2):377-385.
  • 9Heine H G,Stevens M P.A capripoxvirus detection PCR and antibody ELISA based on the major P32,the homolog of the vaccinia virus H3L gene[J].J Immun Methods,1999,227:187-196.
  • 10Carn V M.Use of a recombinant antigen in an indirect ELISA for detecting bovine antibody to capripoxvirus[J].J Immun Methods,1994,49:285-294.

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7

引证文献1

二级引证文献3

中国兽医学报
2009年 第8期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部