深圳大学学报理工版

曹会,蔡德丰,杨平常,等.家蚕MBP-Bmlp7蛋白的克隆表达及免疫学鉴定[J].深圳大学学报理工版,2017,34(No.2(111-220)):117-121.[doi:10.3724/SP.J.1249.2017.02117]
Cao Hui,Cai Defeng,Yang Pingchang,et al.Expression and immunological identification of MBP-Bmlp7 from Bombyx mori[J].Journal of Shenzhen University Science and Engineering,2017,34(No.2(111-220)):117-121.[doi:10.3724/SP.J.1249.2017.02117]

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家蚕MBP-Bmlp7蛋白的克隆表达及免疫学鉴定

曹会¹,蔡德丰²,杨平常¹,刘志刚¹,夏立新¹

1)深圳大学医学部,呼吸疾病国家重点实验室深圳大学变态反应分室,广东深圳518060; 2)深圳市儿童医院检验科,广东深圳 518026

关键词：免疫学; 家蚕; 30 kD脂蛋白; 表达纯化; 免疫印迹; 融合蛋白; 免疫原性

Expression and immunological identification of MBP-Bmlp7 from Bombyx mori

Cao Hui¹, Cai Defeng², Yang Pingchang¹, Liu Zhigang¹, and Xia Lixin^1

Cao Hui¹, Cai Defeng², Yang Pingchang¹, Liu Zhigang¹, and Xia Lixin^11)Health Science Center, State Key Laboratory of Respiratory Disease for Allergy at Shenzhen University, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China 2)Department of Clinical Laboratory, Shenzhen Children's Hospital, Shenzhen 518026, Guangdong Province, P.R.China

Keywords： immunology; Bombyx mori; 30 kD lipoprotein; expression and purification; immunoblotting; fusion protein; immnogenicity

DOI: 10.3724/SP.J.1249.2017.02117

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参考文献

克隆表达家蚕蚕蛹30 kD(1 kD=1 u)脂蛋白家族成员Bmlp7(Bombyx mori lipoprotein 7)的融合蛋白,鉴定其致敏原性.合成Bmlp7的基因后连接至载体质粒pMAL-C5e上,在大肠杆菌BL21中诱导表达出麦芽糖结合蛋白(maltose binding protein, MBP)-Bmlp7,纯化后,利用Western blot鉴定MBP-Bmlp7融合蛋白与家蚕过敏患者血清中特异性免疫球蛋白E(immunoglobulins E, IgE)的结合能力.结果显示,纯化得到了纯度较高的MBP-Bmlp7融合蛋白,且该融合蛋白能够与家蚕过敏患者血清特异性IgE结合,说明Bmlp7是家蚕中一种潜在的致敏原.

The open reading frame of Bmlp7 from silkworm(Bombyx mori)was chemically synthesized and cloned into plasmid pMAL-c5e. The recombinant plasmid pMAL-Bmlp7 was transformed into E.coli BL21 and expressed as a fusion protein. The protein was purified using amylose affinity chromatography and purified to homogeneity. The immunoglobulins E(IgE)reactivity of the fusion protein was assayed by western blotting using sera of patient with silkworm allergies. The results show that the Bmlp7 fusion protein has IgE reactivity with the patient sera.

引言
1 材料与方法
2 实验结果
3 讨论

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