Gene cloning and expression purification of human autoimmune antigen SSA/60 / 国际检验医学杂志

ABSTRACT

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 × 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 .