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抗禽流感病毒H5N1分泌型IgA在中国仓鼠卵巢细胞中的表达 被引量:1

Expression of anti-avian influenza virus H5N1 secretory IgA in Chinese hamster ovary cells
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摘要 分泌型IgA(SIgA)在机体的粘膜免疫中具有重要作用,在外分泌道中比单体IgA和IgG抗体具有更好的抗感染活性。为了表达抗禽流感病毒H5N1人-鼠嵌合分泌型IgA抗体,首先以本室先前构建的稳定表达IgA的中国仓鼠卵巢细胞(CHO)细胞系为基础,共转染分泌片和J链表达质粒,然后用抗生素Zeocin选择阳性克隆细胞,利用倍比稀释的方法筛选分泌SIgA的单克隆细胞,通过Western blotting分析培养上清中SIgA的表达情况。结果表明,在CHO细胞中成功表达了SIgA抗体,上述研究为研制分泌型IgA抗体制剂奠定了良好的基础。 Secretory IgA(SIgA) antibodies in external secretions play an important role in mucosal immune response.Polymeric SIgA was advantageous over monomeric IgA(mIgA) and IgG in several aspects.To express secretory IgA antibody against H5N1 virus,we constructed the secretory component and immunoglobulin J expressing plasmids and co-transfected the plasmids into the Chinese hamster ovary cells(CHO) stably expressing immunoglobulin A.Then we used Zeocin to select the positive clone cells,monoclonal cells stably secreting SIgA was screened through fold dilution method at last.The SIgA antibody secreted from the CHO cells was confirmed by Western blotting,which demonstrated that we had got the complete SIgA molecular.The successful expression of this polymeric anti-H5N1 SIgA in CHO cells will contribute to the production of recombinant SIgA as a preventive agent for infectious disease control.
出处 《生物工程学报》 CAS CSCD 北大核心 2011年第2期219-225,共7页 Chinese Journal of Biotechnology
基金 国家自然科学基金(No.30872223) 国家高技术研究发展计划(863计划)(No.2009AA02Z111)资助~~
关键词 禽流感病毒H5N1 WESTERN BLOTTING SIgA抗体 Avian influenza virus H5N1 Western blotting secretory Immunoglobulin A
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参考文献10

  • 1WHO. Cumulative Number of Confirmed Human Cases of Avian Influenza A/(H5N1) Reported to WHO. http://www.who.int/csr/disease/avian_influenza/country/cases_table_2010_03_29/en/index.html, 2010.
  • 2Corthesy B. Recombinant immunoglobulin A: powerful tools for fundamental and applied research. Trends Biotechnol, 2002, 20(2): 65-71.
  • 3Ma JK, Hikmat BY, Wycoff K, et al. Characterization of a recombinant plant monoclonal secretory antibody and preventive immunotherapy in humans. Nat Med, 1998, 4(5): 601-606.
  • 4Dallas SD, Rolfe RD. Binding of Clostridium difficile toxin A to human milk secretory component. J Med Microbiol, 1998, 47(10): 879-888.
  • 5Phalipon A, Cardona A, Kraehenbuhl JP, et al. Secretory component: a new role in secretory IgA-mediated immune exclusion in vivo. Immunity, 2002, 17(1): 107-115.
  • 6张宝中,安小平,张昕,刘大斌,单云竹,周育森,童贻刚.用基因组DNA剪接技术克隆SIgA相关基因[J].中国生物工程杂志,2008,28(6):1-6. 被引量:2
  • 7张宝中,冉多良,张昕,安小平,单云竹,周育森,童贻刚.用DREAM技术进行全长质粒快速定点突变[J].生物工程学报,2009,25(2):306-312. 被引量:3
  • 8张宝中,张昕,陈万荣,刘大斌,王盛,安小平,冉多良,赵光宇,周育森,童贻刚.抗H5N1病毒嵌合IgA抗体基因的构建及其在CHO细胞中的表达[J].生物工程学报,2009,25(5):714-719. 被引量:1
  • 9Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. New York: Cold Spring Harbor Laboratory Press, 1989: 880-897.
  • 10Hammarstrom L, Weiner CK. Targeted antibodies in dairy-based products. Adv Exp Med Biol, 2008, 606: 321-343.

二级参考文献38

  • 1朱晓峰,安小平,陈锦辉,陈士华,张昕,周育森,童贻刚.一种简便、高效、经济的从凝胶中回收DNA的方法[J].生物技术通讯,2006,17(4):603-604. 被引量:14
  • 2朱晓峰,陈锦辉,张昕,安小平,周育森,童贻刚.用DREAM技术纠正人工合成基因中的突变[J].中国生物工程杂志,2007,27(1):86-92. 被引量:2
  • 3Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: A Laboratory Manual. 3nd ed. New York: Cold Spring Harbor Laboratory Press, 2001.
  • 4Aiyar A, Xiang Y, Leis J. Site-directed mutagenesis using overlap extension PCR. Methods Mol Biol, 1996, 57(2): 177-191.
  • 5Kunkel TA. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA, 1985, 82(2): 488-492.
  • 6Weiner MP, Costa GL, Schoettlin W, et al. Site-directed mutagenesis of double-stranded DNA by the polymerase chain reaction. Gene, 1994, 151(1/2): 119-123.
  • 7Li S, Wilkinson MF. Site-directed mutagenesis: a two-step method using PCR and DpnI. Biotechniques, 1997, 23(4): 588-590.
  • 8Wei D, Li M, Zhang X, Xing L. An improvement of the site-directed mutagenesis method by combination of megaprimer, one-side PCR and DpnI treatment. Anal Biochem, 2004, 331(2): 401-403.
  • 9Shenoy AR, Visweswariah SS. Site-directed mutagenesis using a single mutagenic oligonucleotide and DpnI digestion of template DNA. Anal Biochem, 2003, 319(2): 335-336.
  • 10Zhu L. In vitro site-directed mutagenesis using the unique restriction site elimination (USE) method. Methods Mol Biol, 1996, 57(1): 13-29.

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