酸性β-半乳糖苷酶基因密码子优化及高效表达  

Optimization of Acid Lac Z and Its Efficient Expression in Pichia PastorisVector

作　　者：冷春玲[1] 

机构地区：[1]辽东学院农学院,辽宁丹东118003

出　　处：《辽东学院学报（自然科学版）》2012年第3期153-156,共4页Journal of Eastern Liaoning University:Natural Science Edition

摘　　要：选用酵母偏好密码子,设计合成一种酸性β-半乳糖苷酶基因,并研究其在毕赤酵母中的高效表达。优化后,密码子适应指数(CAI)由原来的0.61提升到0.85。在基因的5'端融合编码α信号肽序列,在3'端融合编码6个组氨酸标签序列,并在N端编码α信号肽和编码成熟酸性β半乳糖苷酶基因间分别引入编码kex2和Ste13裂解信号序列。基因克隆到pPICZα-A表达载体,构建成分泌型重组酵母表达载体pPICZα-β-Gal。在醇氧化酶(AOX1)启动子调控下,酸性β-半乳糖苷酶得到高效分泌表达,达到508 mg/L,比优化前提高3倍。重组酸性β-半乳糖苷酶具有天然的酸性β半乳糖苷酶相同的酶活性。An acid Lac Z was designed by using yeast preference codon to study its expression in Pichia Pasto- ris Vector. After optimizing, the codon adaptation index （CAI） rose from 0.61 to 0.85. In this experiment, we linked fusion coding α signal peptide sequence to 5＇ - end of the gene, and linked six His tag sequences to 3＇ - end of the gene. In addition, at N - end, kex2 and Stel3 cracking signal sequences were inserted between ot signal peptide sequence and acid Lac Z gene. This gene was cloned to pPICZot - A expression vector to be a secretory re- structuring yeast expression vector. Under regulate and control of alcohol oxidase （AOX1） actosidase is obtained high expression to 508 mg/L which is three times than before. This galactosidase owns the same enzymatic activity as the nature one promoter, acid β - gal- recombination acid β -galactosidase owns the same enzymatic activity as the nature one.

关 键 词：酸性β-半乳糖苷酶 密码子适应指数 毕赤酵母 分泌表达 

分 类 号：Q753[生物学—分子生物学]