[1]邬玉兰,刘志刚.大豆主要过敏原Gly m Bd 30K基因的克隆及其原核表达载体的构建[J].大豆科学,2009,28(01):11-15.[doi:10.11861/j.issn.1000-9841.2009.01.0011]
　WU Yu-lan,LIU Zhi-gang.Cloning and Prokaryotic Expression Vector Construction of Gly m Bd 30K Gene from Soybean(Glycine max)[J].Soybean Science,2009,28(01):11-15.[doi:10.11861/j.issn.1000-9841.2009.01.0011]
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大豆主要过敏原Gly m Bd 30K基因的克隆及其原核表达载体的构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第28卷 期数: 2009年01期 页码: 11-15 栏目: 出版日期: 2009-02-25
Title:
Cloning and Prokaryotic Expression Vector Construction of Gly m Bd 30K Gene from Soybean(Glycine max)
文章编号:
1000-9841(2009)01-0011-05
作者:
邬玉兰; 刘志刚
深圳大学过敏反应与免疫学研究所，广东 深圳 518060
Author(s):
WU Yu-lan; LIU Zhi-gang
Allergy and Immunology Institute，Shenzhen University，Shenzhen 518060，Guangdong,China
关键词:
大豆; 过敏原; Gly m Bd 30K; 克隆; 原核表达载体
Keywords:
Soybean(Glycine max); Allergen; Gly m Bd 30K; Cloning; Prokaryotic Expression Vector
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2009.01.0011
文献标志码:
A
摘要:
利用RT-PCR克隆大豆主要过敏原Gly m Bd 30K的全长基因，根据序列设计带有酶切位点的特异性引物，扩增大豆Gly m Bd 30K的完整开放阅读框，与pET-28a载体连接，构建原核表达载体。结果表明：克隆了大豆主要过敏原Gly m Bd 30K基因，且构建了其原核表达载体。该基因含有长度为1 140 bp的开放阅读框，编码379个氨基酸。该蛋白质的相对分子质量为42 758，等电点为5.08。序列同源性分析发现其与数据库中已知的Gly m Bd 30K基因同源性很高，因此认为其是大豆的过敏原基因，在GenBank数据库中的登录号为EU883600。克隆的大豆主要过敏原Gly m Bd 30K基因及构建的原核表达载体，为大豆主要过敏原Gly m Bd 30K的重组表达和免疫活性鉴定等奠定基础。
Abstract:
The RT-PCR was applied to clone the full- length allergen genes from soybean and the sequences were analyzed.The specific primers were designed.The ORF of Gly m Bd 30K of soybean was subcloned into the expression vector pET 28a.Results showed that the open reading frame（ORF）of cloned cDNA contained 1 140 bp and encoded 379 amino acids，and the estimated molecular mass of the encoded protein was 42 758 with pI 5.08.Sequence analysis showed that this clone shared high identities with Gly m Bd 30K from soybean.The deduced protein was therefore regarded as the major allergen of Gly m Bd 30K of soybean（GenBank database entry No.EU883600）.Cloning and prokaryotic Expression Vector Construction of Gly m Bd 30K gene from soybean，which will be used as base for the expression and identification of allergen protein.

参考文献/References:

[1] WHO Workshop.Asthma management and prevention[R].The global initiation for asthma ascretariate，1996：1-2.

[2] Metcalfe D.The nature and mechanisms of food allergies and related diseases[J].Food Technology，1992，5（5）：136-140.
[3] Herian A M，Taylor S L，Bush R K.Identification of soybean allergens by immunoblotting with sera from soy-allergic adults[J].International archives of allergy and applied immunology，1990，92（2）：193-198.
[4] Vidal C，Perez-Carral C，Chomon B.Unsuspected sources of soybean exposure [J].Annals of Allergy，Asthma & Immunology，1997，79（4）：350-352.
[5] Kalinski A J，Melroy D L，Dwivedi R S，et al.A soybean vacuolar protein（P34）related to thiol proteases which is synthesized as a glycoprotein precursor during seed maturation[J].Journal of Biological Chemistry，1992，15；267(17)：12068-12076.
[6] Kalinski A J，Weisemann J，Matthews B F，et al.Molecular cloning of a protein associated with soybean oil bodies which is homologous to thiol proteases of the papain family[J].Journal of Biological Chemistry，1990，265(23)：13843-13848.
[7] Ogawa T，Tsuji H，Kitamura K，et al.Identification of the soybean allergenic protein，Gly m Bd 30 K，with the soybean seed 34-kDa oil-body-associated protein[J].Bioscience，Biotechnology，and Biochemistry.1993，57(6)：1030-1033.
[8] Vrtala S，Focke-Tejkl M，Swoboda I，et al.Strategies for converting allergens into hypoallergenic vaccine candidates [J].Methods，2004，32(3)：313-20.
[9]吕相征，刘秀梅.健康人群食物过敏状况的初步调查[J].中国食品卫生杂志.2005，17(2)：119-120.（Lu X Z，Liu X M.Health status of food allergy crowd preliminary investigation[J].Chinese Journal of Food Hygiene，2005，17(2)：119-120.）
[10]郑云兰，李霞辉.大豆营养分析技术[M].哈尔滨：黑龙江科学技术出版社，1991：190.(Zheng Y L，Li X H.Soybean nutritional analysis[M].Harbin：Heilongjiang Science and Technology Publishing House，1991：190.)
[11]李钢，丁淑霞，左容，等.食物过敏引起变态反应疾病184例临床分析[J].佳木斯医学院学报，1989，12（3）：269.(Li G，Ding S X，Zuo R，et al.Food allergies cause allergic disease clinical analysis of 184 cases[J].Jiamusi Medical College，1989，12（3）：269.)
[12] Duke W W.Soybean as a possible important source of allergy[J].Allergy，1934(5)：300-304.
[13] Castimpoolas N.Isolation of a-，β-，and γ-conglycinins[J].Archives of Biochemistry and Biophysics，1969，129：409-497.
[14] Ogawa T，Bando N，Tsuji H，et al.Investigation of the IgE binding proteins in soybeans by immunoblotting with the sera of the soybean sensitive patient s with atopic dermatitis[J].Journal of Nutritional Science and Vitaminology，1991，37（6）：555-565.
[15] Bando N,Tsuji H,Yamanishi R，et al.Identification of the glycosylation site of a major soybean allergen，Gly m Bd 30K[J].Bioscience，Biotechnology，and Biochemistry 1996，60(2)：347-348.
[16] Rost B.Twilight zone of protein sequence alignments [J].Protein Engineering，1999，12：85-94.

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备注/Memo

基金项目：广东省科技重点专项资助项目(2003A3080502)，深圳市科技计划资助项目（200326）。

作者简介：邬玉兰(1988-)，女，实验员，主要从事过敏原的分子生物学研究。E-mail:xinyi880826@163.com。

更新日期/Last Update: 2014-09-29