Abstract:

Objective To construct the prokaryotic expression vector pET28a-YARA-Ihc-ProDer f 1 and provide the foundation for exploring the fusion protein effect as vaccine for specific immunotherapy. Methods Two oligonucleotides encoding YARA were synthesized and annealed to generate YARA- encoding DNA. The fused genes, YARA- Ihc- ProDer f 1 was constructed and inserted into the prokaryotic expression vector pET28a(+). The fusion protein YARA-Ihc-ProDer f 1 induced with IPTG in E.coli BL21(DE3) was purified with Ni2 +-resin affinity chromatography and confirmed with SDS-PAGE and Western blot. Results Sequence analysis confirmed the construction of the expression vector pEt28a- YARA-Ihc-ProDer f 1 successfully, the fusion protein YARA-Ihc-ProDer f 1 was expressed and purified with the concentration of 278 μg/ml. SDS-PAGE and Western blot demonstrated the fusion protein was YARA-Ihc-ProDer f 1. Conclusion
The recombinant prokaryotic expression vectors, pET28a(+ )- YARA- Ihc- ProDer f 1 was successfully constructed, and the fusion protein was expressed and purified .

Key words: Cell penetrating peptides, Dermatophagoides farina, invariant chain, allergens