期刊文献+

鸡传染性法氏囊病毒强毒株(vvIBDV)VP_2高变区的替换

下载PDF
导出
摘要 采用双融合PCR(doublefusionPCR)的方法将鸡传染性法氏囊病毒 (IBDV)的强毒株的VP2 片段替换为疫苗株的相应片段以获得重组的病毒粒子。首先将被替换片段两侧的片段从强毒株的载体上扩增下来 ,并将目的片段从疫苗株序列的载体上扩增下来。然后用融合PCR的方法将目的片段与其上游片段融合起来 ,回收产物并将其与下游片段融合起来 ,最终得到了替换后的新序列。双融合PCR提供了一种快速、精确和不受酶切位点限制的片段替换方法。
作者 苗靳
出处 《山西医科大学学报》 CAS 2002年第6期500-502,共3页 Journal of Shanxi Medical University
  • 相关文献

参考文献7

  • 1[1]Brandt M, Yao K, Liu M, et al. Molecular determinants of virulence, cell tropism and pathogenic phenotype of infectious bursal disease virus[J]. J Virol, 2001,75:11974~11982.
  • 2[2]Van Loon AA, de Haas M, Zeyda I, et al. Alternation of amino acids in VP2 of very virulent infectious bursal disease virus results in tissue culture adaptation and attenuation in chickens[J] .J Gen Virol, 2002, 83:121~129.
  • 3[3]Amberg DC, Botstein D, Beasley EM.Precise gene disruption in Saccharomyces cerevisiae by double fusion polymerase chain reaction[J]. Yeast, 1995,11:1275~1280.
  • 4[4]Horton RM, Hunt HD, Ho SN, et al. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension[J]. Gene, 1989, 77: 61~68.
  • 5[5]Ho SN, Hunt HD, Horton RM, et al. Site-directed mutagenesis byoverlap extension using the polymerase chain reaction[J]. Gene, 1989, 77:51~59.
  • 6[6]Jespersen T, Duch M, Pedersen FS. Efficient non - PCR - mediated overlap extension of PCR fragments by exonuclease"End Polishing" [J]. Biotechniques, 1997, 23: 48~52.
  • 7[7]Warrens AN, Jones MD, Lechler RI. Splicing by overlap extension by PCR using asymmetric amplification: an improved technique for the generation of hybrid proteins of immunological interest [J] .Gene, 1997, 186:29~35.

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