花生过敏原Ara h2.02原核表达方法条件的研究

被引：3

作者：

蔡琴

张文举

陈沁

机构：

[1] 上海大学生命科学学院

来源：

食品与机械 | 2015年 / 31卷 / 02期

关键词：

花生; 过敏原; 重组蛋白; Ara h 2.02; 原核表达;

D O I：

10.13652/j.issn.1003-5788.2015.02.009

中图分类号：

TS201.2 [食品化学]; R446.6 [免疫学检验];

学科分类号：

083201 ; 100208 ;

摘要：

将重组质粒pET-32a(+)-Arah2.02转化表达宿主菌Rosetta(DE3)中,经IPTG诱导表达,SDS—PAGE电泳分析,结果显示表达的蛋白大小约为38kDa。进一步用通用His标签抗体进行Western Blotting检测,结果表明成功克隆表达了花生过敏原Arah2.02。为获得较多的重组蛋白Arah2.02,分别对IPTG浓度、摇床转速、诱导温度和时间等条件进行选择,确定最佳条件为:IPTG浓度0.3mmol/L,摇床转速220r/min,诱导温度37℃,诱导时间2h。

引用

页码：43 / 46+156 +156

页数：5

共 22 条

[1]

花生过敏原蛋白Ara h1的纯化表达研究.[D].闫飞.南昌大学.2012, 03

[2] Food allergy: Epidemiology, pathogenesis, diagnosis, and treatment [J].

Sicherer, Scott H. ;

Sampson, Hugh A. .

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 2014, 133 (02) :291-+

[3] IgE cross-reactivity between the major peanut allergen Ara h 2 and the nonhomologous allergens Ara h 1 and Ara h 3 [J].

Bublin, Merima ;

Kostadinova, Maria ;

Radauer, Christian ;

Hafner, Christine ;

Szepfalusi, Zsolt ;

Varga, Eva-Maria ;

Maleki, Soheila J. ;

Hoffmann-Sommergruber, Karin ;

Breiteneder, Heimo .

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY, 2013, 132 (01) :118-U212

[4]

Evidencing the role of lactose permease in IPTG uptake by Escherichia coli in fed-batch high cell density cultures.[J].Alfred Fernández-Castané;Claire E. Vine;Glòria Caminal;Josep López-Santín.Journal of Biotechnology.2011, 3

[5] Primary sequence and site-selective hydroxylation of prolines in isoforms of a major peanut allergen protein Ara h 2 [J].

Li, Jinxi ;

Shefcheck, Kevin ;

Callahan, John ;

Fenselau, Catherine .

PROTEIN SCIENCE, 2010, 19 (01) :174-182

[6] Awake intubation with the Bonfils fibrescope in patients with difficult airway [J].

Corbanese, Ulisse ;

Possamai, Clemente .

EUROPEAN JOURNAL OF ANAESTHESIOLOGY, 2009, 26 (10) :837-841

[7]

Crystal structure of Ara h 3; a major allergen in peanut.[J].Tengchuan Jin;Feng Guo;Yu-wei Chen;Andrew Howard;Yu-Zhu Zhang.Molecular Immunology.2009, 8

[8] Peanut allergy [J].