Cloning and expression of nogo-66 protein in prokaryotic cells

Objective: To clone Nogo-66 gene,construct the recombinant prokaryotic expressive vector and express its products in E.coli.Methods: Nogo-66 gene was amplified by reverse transcription polymerase chain reaction(RT-PCR) and then cloned into PMD18-T vector.After identified by the enzyme digestion,the gene was subcloned into prokaryotic expressive vector pET-42a(+),which was identified by the enzyme digestion and sequencing.Nogo-66 fusion protein was expressed in E.coli BL21(DE3) by IPTG.Results: The Nogo-66 gene and recombinant prokaryotic expression vector were obtained and the Nogo-66 protein was expressed and increased with prolong of time.Conclusions: The Nogo-66 expression vector and its prokaryotic expression product were obtained.It maybe has a great significance for study the biologic function of Nogo-66 protein and the preparation of monoclonal antibody against Nogo-66 protein.