密码子优化的重组黄曲霉毒素解毒酶（rADTZ）在毕氏酵母中组成型分泌表达的研究

›› 2007, Vol. 9 ›› Issue (5): 87-94.

密码子优化的重组黄曲霉毒素解毒酶（rADTZ）在毕氏酵母中组成型分泌表达的研究

左振宇刘大岭胡亚冬胡熔姚冬生

暨南大学（科仁）微生物技术研究所,广州510632

收稿日期:2007-06-21 修回日期:2007-09-01 出版日期:2007-10-15 发布日期:2009-10-19
通讯作者: 刘大岭，教授，研究方向为应用微生物学。Tel：020—85228422；E-mail：tldl@jnu．edu．cn
作者简介:左振宇|硕士研究生|从事分子生物学和微生物学研究。
基金资助:
国家十五“863”计划项目“黄曲霉毒素解毒酶基因的克隆及其基因工程菌的构建”（2002AA213011;2005AA213010）和国家自然科学基金项目“黄曲霉毒素解毒酶基因的克隆及表达的研究”（30270043）资助.

Studies on Constitutive and Secretive Expression of Codon-Optimized |Recombinant Aflatoxin-Detoxifizyme （rADTZ） in Pichia pastoris

ZU0 Zhen-yu, LIU Da-ling, HU Ya-dong, HU Rong, YA0 Dong-sheng

Institute of Microbial Biotechnology, Jinan University, Guangzhou 510632, China

Received:2007-06-21 Revised:2007-09-01 Online:2007-10-15 Published:2009-10-19

摘要/Abstract

摘要：

黄曲霉毒素解毒酶（ADTZ）来源于真菌Armillariellatabescens，它能够有效地分解黄曲霉毒素。为了使重组蛋白rADTZ能在毕氏酵母中高效分泌表达，根据毕氏酵母密码子的偏好性对rADTZ的5’末端的编码区域进行了优化，利用two—stepDNAsynthsis技术合成出ADTZ优化的基因序列OPT—ADTZ，并与组成型表达载体pGAPZaA连接，构建重组质粒pNOA，线性化pNOA后转化至毕氏酵母GS115中，实现了密码子优化的rADTZ组成型分泌表达。

关键词: ADTZ two—step DNA synthesis 组成型分泌表达

Abstract:

Aflatoxin-detoxifizyme （ADTZ） , being from Armillariella tabescens, can effectively decompose aflatoxins. The 5＇coding region of its eDNA was optimized according to the codon bias of Pichia pastoris to make it secretively express in Pichia pastoris with higher performance. Two-step DNA synthsis was used to synthesize the eDNA sequence being optimized of ADTZ （OPT-ADTZ）. OPT-ADTZ was connected with constitutive plasmid pGAPZaA to construct the reecombinant plasmid pNOA. pNOA was linearized and then transformed into Pichia pastoris GS115. Then codonoptimized ADTZ was constitutively and secretively expressed in Pichia pastoris.

Key words: ADTZ , two-step DNA synthesis, constitutive and secretive expression

中图分类号:

Q78 Q814

左振宇刘大岭胡亚冬胡熔姚冬生. 密码子优化的重组黄曲霉毒素解毒酶（rADTZ）在毕氏酵母中组成型分泌表达的研究[J]. , 2007, 9(5): 87-94.

ZU0 Zhen-yu, LIU Da-ling, HU Ya-dong, HU Rong, YA0 Dong-sheng. Studies on Constitutive and Secretive Expression of Codon-Optimized |Recombinant Aflatoxin-Detoxifizyme （rADTZ） in Pichia pastoris[J]. , 2007, 9(5): 87-94.