The Codon Optimization Of ADTZ And Construction Of Combinant PPNOA

Aflatoxins are potent hepatocarcinogen. The aflatoxin B1 of them is the most potent toxic. It is a hot topic that the detoxification of aflatoxins in this studying field. The fungal E-20, that produce aflatoxin-detoxifizyme, has been screened by our research team. At present we have successfully got the full length cDNA named E123 of aflatoxin-detoxifizyme and complete ORF sequence named ADTZ by the technology of gene engineering.In order to high-level expression of aflatoxin-detoxifizyme, Combinant pSA has been successfully obtained and ADTZ by our research institution. ADTZ has been expressed extracellularly in Pichia.Pastoris,but the level of ADTZ expressed is't high enough to advanced research and let alone industrial production. In this article, we are trying to Optimize the ADTZ according to the codon bias of Pichia.Pastoris, hoping that the ADTZ could be expressed with the high level in PPastorisWe divided the optimization 5' end of ADTZ' spaned 936bp into five fraction named individually Opt-p3, Opt-A1, Opt-A2, Opt-A3, Opt-A4. The length corresponding to each fraction was 219bp, 270bp, 192bp, 183bp, 159bp。 With the help of Two-step DNA synthesis method, we synthesized this five fraction firstly, and then made them into a complete sequence named Opt-A'. At last splicing Opt-A' and non- optimization ADTZ-B, we obtained the complete ORF sequence named N-Opt-ADTZ.Combinant pNOA was constructed by inserting the N-Opt-ADTZ into pPICzaA for the advanced research in P.Pastoris.