大豆主要过敏原β-伴大豆球蛋白α亚基的分段及克隆载体的构建

Overlapping segments and cloning vector construction of α-subunit of β-conglycinin, the major soybean allergen

利用生物信息学软件将大豆主要过敏原β-伴大豆球蛋白α亚基分为相互重叠的4个片段(A、B、C、D),设计和合成系列引物。利用PCR技术分别扩增β-伴大豆球蛋白α亚基Overlapping分段基因,将其插入到p MD-18T vector中,对重组质粒进行PCR和双酶切鉴定,测序结果与设计基因序列一致,成功构建了克隆载体。为研究β-伴大豆球蛋白α亚基的分段表达及抗原表位的筛选提供参考。

The α-subunit of β-conglycinin was divided into four overlapping fragments segmented （A, B, C and D） by bioinformatics tool, and a series of primers were designed and synthesized. The overlapping α-subunit of β-conglycinin genes were amplified by PCR and inserted into pMD-18T vector. The recombinant plasmids was evaluated by PCR and double enzyme digestion, and DNA sequence analysis showed that the cloned sequence was the very fragment designed. The successful cloning of the genes could provide a reference for the study of the expression of α subunit of β-conglycinin and the screening of antigenic epitopes.