Cloning and Expression of the Antigenic Epitope of Gly m Bd 30K Protein from Soybean and Purification and Identification of Expressed Product(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:

2010年02期

Page:

186-190

Research Field:

Publishing date:

Info

Title:

Cloning and Expression of the Antigenic Epitope of Gly m Bd 30K Protein from Soybean and Purification and Identification of Expressed Product

Author(s):

LIN Su-xia¹; 2;  WANG Xiao-mei³;  LIU Zhi-gang¹; 3;  ZENG Meng-ya¹; WU Yan¹; CHEN Jia-jie¹

1.State Key Laboratory of Respiratory Disease for Allergy, Shenzhen University, Shenzhen 518060;

2.College of Life Science , Shenzhen University, Shenzhen 518060;

3.Medical College, Shenzhen University, Shenzhen 518060, Guangdong, China

Keywords:

The major allergens of soybean;  Gly m Bd 30k;  Epitope;  Cloning

PACS:

S565.1

DOI:

10.11861/j.issn.1000-9841.2010.02.0186

Abstract:

Through reviewing the literatures combined with bioinformatics predication, the antigenic epitope of Gly m Bd 30k allergen was selected. And the gene was amplified by PCR. The pET expression vector for the monomer protein (pET-sGly) and the dimer (pET-dGly) were constructed. The recombinant proteins were expressed in E.coli BL21 (DE3) plysS by IPTG, analyzed by SDS-PAGE, and then purified by metal (Ni2+) chelating affinity chromatography.The immunogenicity was tested by western-blotting and ELISA. Results shows that the monomer protein sGly was mainly expressed as soluble protein, while the dimer protein dGly as inclusion bodies. Purified protein sGly and dGly both have good immunogenicity, and sGly is better. The prokaryotic expression vectors for the monomer protein and dimer of the antigenic epitope of Gly m Bd 30k protein of soybean will make great foundations for developing the monoclonal antibodies and the detection kit of the major allergens of soybean.

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