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Expression, purification and biological activity analysis of the epitope peptide of human tyrosinase antigen
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF DERMATOLOGY
- Issue:
- 3
- DOI:
- 10.3760/j.issn:0412-4030.2007.03.003
- Key Word:
- 白癜风;一元酚单氧酶;原核表达
Abstract： 目的 探讨人酪氨酸酶抗原表位肽在白癜风患者自身抗体检测中的表达.方法 目的 基因克隆至原核表达载体pGEX-4T-2,转化大肠杆菌BL21菌株,异丙基-β-D-硫代半乳糖苷(IPTG)诱导蛋白表达,通过SDS-PAGE分析和蛋白质印迹鉴定目的 蛋白表达,ELISA检测其生物学活性.结果 成功构建重组表达载体,SDS-PAGE和蛋白质印迹结果表明,重组蛋白成功表达.应用凝胶分析系统分析蛋白表达量发现,目的蛋白的表达量占菌体总蛋白的90%.融合蛋白经过试剂盒纯化后,其蛋白纯度达95%以上.ELISA检测纯化蛋白的生物学活性,结果表明,纯化蛋白具有结合白癜风患者IgG的能力.结论 成功表达了人酪氨酸酶抗原表位肽,其表达蛋白具有生物学活性.
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