经合理的序列重组制备花生主要过敏原Ara h 2低致敏衍生物

被引：4

作者：

易海涛 ^{[1
,2
]}

刘芳 ^{[3
]}

夏立新 ^{[2
,4
]}

闫浩 ^{[1
,2
]}

刘飞燕 ^{[2
,4
]}

李建杰 ^{[1
,2
]}

刘志刚 ^{[2
,4
]}

机构：

[1] 深圳大学生命科学学院

[2] 深圳大学医学院过敏反应与免疫学研究所

[3] 新疆华世丹药业股份有限公司

[4] 深圳大学医学院

来源：

中国生物工程杂志 | 2011年 / 31卷 / 07期

关键词：

花生; 重组; 过敏原 Ara h 2; 低致敏衍生物; 疫苗;

D O I：

10.13523/j.cb.20110709

中图分类号：

R392.11 [免疫分子生物学（免疫化学）];

学科分类号：

100102 ;

摘要：

目的:构建经序列重组的Ara h 2表达载体,表达并纯化该蛋白,鉴定其低致敏原性。方法:根据已鉴定的Ara h 2 IgE抗原表位,利用基因工程技术将Ara h 2基因进行合理的组合,并将其序列进行合成,再将合成后的基因连入原核表达载体pET-32a(+)上,然后转入Origami宿主表达菌中;IPTG诱导表达;通过Ni2+亲和层析(FPLC)纯化目的蛋白;Western blotting和ELISA鉴定该重组蛋白的低致敏原性。结果:测序结果表明合成后的序列成功转入原核表达载体pET-32a(+)上。重组蛋白纯化后经SDS-PAGE鉴定,目的蛋白大小与理论值相符。Western blotting和ELISA结果均表明通过基因工程改造的Ara h 2蛋白(S-Ara h 2)与重组的Ara h 2(R-Ara h 2)蛋白相比,结合花生过敏病人混合血清中IgE显著降低。结论:成功构建了经序列重组的Ara h 2表达载体,该基因表达的重组蛋白具有良好的低致敏原性,这将会为花生过敏患者的脱敏治疗提供了新的安全疫苗基础。

引用

页码：54 / 59

页数：6

共 24 条

[1]

Prevalence of Peanut and Tree Nut Allergy in the UnitedStates Determined by Means of a Random Digit Dial Tele-phone Survey:a 5-year Follow-up Study. SICHERER S H,MUNOZ-Furlong A,SAMPSON H A. The Journal of Allergy and Clinical Immunology . 2003

[2]

The future of antigen-specific immunotherapy of allergy. Valenta R. Nature Reviews Immunology . 2002

[3]

Allergy or Tolerance in Children Sensitized to Peanut:prevalence and Differentiation Using Component-resolvedDiagnostics. NICOLAOU N,POORAFSHAR M,MURRAY C,et al. The Journal of Allergy and Clinical Immunology . 2010

[4]

Designing Hypoallergenic Derivatives for Allergy Treat-ment by Means of in Silico Mutation and Screening. THALHAMER T,DOBIAS H,STEPANOSKA T,et al. The Journal of Allergy and Clinical Immunology . 2010

[5] Designing hypoallergenic derivatives for allergy treatment by means of in silico mutation and screening [J].

Thalhamer, Theresa ;

Dobias, Heidi ;

Stepanoska, Tatjana ;

Proell, Martina ;

Stutz, Hanno ;

Dissertori, Oliver ;

Lackner, Peter ;

Ferreira, Fatima ;

Wallner, Michael ;

Thalhamer, Josef ;

Hartl, Arnulf .

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 2010, 125 (04) :926-934

[6]

Further fatalitiescaused by anaphylactic reactions to food,2001-2006. Bock S A,Munoz-Furlong A,Sampson H A. The Journal of Allergy and Clinical Immunology . 2007

[7]

Allergen Immunotherapy: Therapeutic Vaccines for Allergic Diseases[J] . J. Bousquet,R. Lockey,H.J. Malling,E. Alvarez-Cuesta,G.W. Canonica,M.D. Chapman,P.J. Creticos,J.M. Dayer,S.R. Durham,P. Demoly,R.J. Goldstein,T. Ishikawa,K. Ito,D. Kraft,P.H. Lambert,H. Lowenstein,U. Muller,P.S. Norman,R.E. Reisman,R. Valenta,E. Valovirta,H. Yssel. &nbspAnnals of Allergy, Asthma & Immunology . 1998 (5)

[8]

Prevalence of peanut and tree nut allergy in the United States determined by means of a random digit dial telephone survey[J] . &nbspThe Journal of Allergy and Clinical Immunology . 2003 (6)

[9] The natural history of peanut and tree nut allergy [J].

Fleischer, David M. .

CURRENT ALLERGY AND ASTHMA REPORTS, 2007, 7 (03) :175-181

[10] 花生致敏原的研究进展 [J].

王通 ;

梁炫强 ;

李玲 .

中国油料作物学报, 2007, (03) :353-358

← 1 2 3 →