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HIV-1跨膜蛋白gp41重组抗原的表达及其免疫反应性 被引量:5

HIV-1 transmembrane glycoprotein 41:Expression of recombinant antigen and antigenicity analysis
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摘要 目的为开发和建立敏感、特异的抗HIV-1抗体的检测方法研制重组gp41抗原。方法根据HIV-1的基因序列,设计并合成了1对PCR扩增引物,应用PCR技术从HIV-1外膜基因组中扩增出HIV-1的gp41截短体,将扩增的基因片段插入质粒pET-28a中构建成重组表达质粒pET-gp41。诱导表达并纯化gp41重组抗原,对gp41进行了初步应用分析。结果gp41在大肠埃希菌BL21(DE3)中获得高表达,表达量占菌体总蛋白量的26.08%。纯化后截短体gp41的纯度为97.94%。经间接ELISA和免疫印迹检测,纯化后的表达产物gp41具有很高的抗原特异性和免疫反应性。结论研制的重组抗原gp41有较强的抗原性和潜在的应用价值。 Objective To develop and establish a new sensitive, specific method for serologic detection of HIV-1. Methods A pair of primer for PCR was designed and synthesized according to the sequence of human immunodefieieney virus type-1 ( HIV-1 ) gene. By using the synthesized primers, the traneated fragment of transmembrane glyeoprotein 41 (gp41) gene was amplified from the envelop-en- coding region in HIV genome by PCR. The amplified DNA fragment was cloned into a plasmid pET-28a and i'eeombinant expression plasmid pET-gp41 was constructed. The recombinant gp41was indueibly expressed and purified. Preliminary applicable analysis for the antigenieity of the recombinant gp41 was conducted. Results The gp41 was highly expressed in E. eoli BL21 ( DE3 ). The amount of gp41 accounted for 26.08% of the total lytie protein. The purity of the purified gp41 was 97.94%. By using ELISA and Western blot, high specificity and strong antigenieity were shown. Conclusion The recombinant gp41 exhibited strong antigenieity, thus its potential availability for HIV diagnosis may be expected.
出处 《临床检验杂志》 CAS CSCD 北大核心 2008年第2期99-101,共3页 Chinese Journal of Clinical Laboratory Science
基金 中国科学院知识创新工程青年人才领域前沿项目资助(0702121YJ1)
关键词 人类免疫缺陷病毒Ⅰ型 gp41抗原 免疫原性 human immunodefieieney virus type 1 glyeoprotein 41 immunogeneeity
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  • 1J萨姆布鲁克 E F 弗里奇 T 曼尼阿蒂斯著 金冬雁 黎孟枫译.分子克隆实验指南 第2版[M].北京:科学出版社,1989..
  • 2Hu D J, Dondero T J, Mastm T D, et al. Global and molecular epidemiology of HIV[M] . Pp. 27-40 In: AIDS and other manifestations of HIV infection. Ed, Wonnser, GP. Lipponcott-Raven Publishers 1998.
  • 3Cart JK, Foley BT, Leitner T, et al. Reference sequences representing the principal genetic diversity of HIV-1 in the pandemic[M] . P Ⅲ-1-Ⅲ-8 In: Human Retrovirus and AIDS 1998 .Eds. Korber B, Foley B, McCutchan F, Mellors J, Hahn B H,Sodroeki J and Kuiken C. Los Alamos National Labondory, Los Alamos, New Mexico.
  • 4路福寿 林文琴.HP在胃十二指肠疾病中的影响[J].中华医学论坛杂志,2003,9(4):23-25.
  • 5Warren JR,Marshall B.Unidentified curved Bacilli on gastric epithelium in active chronic gastritis[J].Lancet,1983,1(4):1273.
  • 6Suganuma M,Kurusu M,Okabe S,et al.Helicobacter py lori membrane protein 1:a new carcinogenic factor of Heli cobacter pylori[J].Cancer Res,2001,61(17):6356.
  • 7Hirota K,Nagata K,Norose Y,et al.Identification of an antigenic epitope in Helicobacter pylori urease that induces neutralizing antibody production[J].Infect Immun,2001,69(11):6597.
  • 8Iizumi T,Yamanishi S,Kumagai Y,et al.Augmentation of Helicobacter pylori urease activity by its specific IgG antibody:implications for bacterial colonization enhancement[J].Biomedical Research,2005,26(1):35.
  • 9de Haan L,Verweij W,Agsteribbe E,et al.The role of ADP-ribosylation and G(M1)-binding activity in the mucosal immunogenicity and adjuvanticity of the Escherichia coli heatlabile enterotoxin and Vibrio cholerae cholera toxin[J].Immunol Cell Biol,1998,76(3):270.
  • 10Tsuji T,Watanabe K,Miyama A.Monomer of the B subunit of heatlabile enterotoxin from enterotoxigenic Escherichia coli has little ability to bind to GM1 ganglioside compared to its coligenoid[J].Microbiol Immunol,1995,39(10):817.

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