摘要
目的 克隆人SSA/Ro60自身抗原并表达纯化,为辅助诊断自身免疫提供物质基础。方法 采用RT-PCR技术扩增SSA/Ro60基因,定向插入pPICZ表达载体,转入毕赤酵母表达系统,将获得的重组蛋白行SDS-PAGE和Western blotting鉴定。结果 扩增出约1.5kb的SSA/Ro60全长序列,获得相对分子质量60×103的重组蛋白,经鉴定具有SSA/Ro60抗原性。结论成功克隆并表达SSA/Ro60,为诊断自身免疫疾病奠定基础。
Objective To clone human autoimmune antigen SSA/Ro60 and to purify its expression to provide the material basis for the assisted diagnosis of human autoimmune diseases .Methods The SSA/Ro60 gene was cloned by RT‐PCR and directionally inserted into expression vector pPICZ .The recombinant plasmid was transformed into Pichia SMD1168 .The obtained recombinant protein was identified by SDS‐PAGE and Western blotting .Results The amplified full‐length sequence was about 1 .5 kb in size . The pPICZ‐SSA positive clone produced a 60 ＆215; 103 recombinant protein which had natural immunogenicity of human autoimmune antigen SSA/Ro60 by SDS‐PAGE and Western blot .Conclusion Human autoimmune antigen SSA/Ro60 is successfully cloned and expressed ,which lays a foundation for diagnosing autoimmune diseases .
出处
《国际检验医学杂志》
CAS
2016年第16期2224-2225,2228,共3页
International Journal of Laboratory Medicine
基金
辽宁省自然科学基金资助项目(02014010058-301)
