大豆致敏原β-伴大豆球蛋白α’亚基的分段与克隆鉴定

Overlapping and cloning identification of α'-subunit β-conglycinin in the soybean allergenic

大豆是引起人们食物过敏的八大过敏原之一,β-伴大豆球蛋白是大豆主要致敏蛋白之一,并且3个亚基α、α’、β都具有致敏性。利用生物信息学软件对β-伴大豆球蛋白α’亚基进行Overlapping(重叠)分段,通过PCR技术扩增到目的片段,与pMD 18-T载体连接;对重组质粒进行PCR和双酶切鉴定,鉴定结果与基因序列一致,成功构建克隆载体。

Soybean is one of the eight allergens that cause people＇s food allergy. β- conglycinin is one of the major allergenic proteins in soybean, and α-subunit, α＇-subunit and β-subunit are both sensitive. The α＇-subunit of β-conglycinin was divided into overlapping fragments by bioinformatics tool. 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 consistent with fragment designed.