药学学报, 2012, 47(10): 1389-1393
引用本文:
杨金玲, 高丽丽, 朱 平, 侯 琦, 王 芬, 于文博, 聂 涛. 蝎毒镇痛活性肽基因BmK AngM1的密码子优化及其真核表达分析[J]. 药学学报, 2012, 47(10): 1389-1393.
YANG Jin-ling, GAO Li-li, ZHU Ping, HOU Qi, WANG Fen, YU Wen-bo, NIE Tao. Codon optimization and eukaryotic expression analysis of the analgesic peptide gene BmK AngM1 from Buthus martensii Karsch[J]. Acta Pharmaceutica Sinica, 2012, 47(10): 1389-1393.

蝎毒镇痛活性肽基因BmK AngM1的密码子优化及其真核表达分析
杨金玲1, 高丽丽1, 朱  平1*, 侯  琦1, 王  芬1, 于文博1, 聂  涛1, 2
(1. 中国医学科学院、北京协和医学院药物研究所, 天然药物活性物质与功能国家重点实验室, 卫生部天然药物生物合成重点实验室, 北京 100050; 2. 河北农业大学现代科技学院, 保定 071000)
摘要:

密码子的偏爱性是影响基因异源表达的重要因素, 通过对外源基因的密码子序列进行优化可提高其表达水平。为了获得蝎毒镇痛活性肽基因BmK AngM1在毕赤酵母中的高效表达, 根据毕赤酵母密码子偏爱性, BmK AngM1基因中的毕赤酵母低利用率密码子突变为其高利用率简并密码子, 克隆到表达载体pPIC9K, 转化毕赤酵母, 甲醇诱导表达; 重组蛋白表达量测定结果显示, 优化后的BmK AngM1基因在毕赤酵母中的表达水平是优化前的3.7倍。研究结果表明, 密码子优化能显著提高BmK AngM1基因在毕赤酵母中的表达水平。

关键词:   
Codon optimization and eukaryotic expression analysis of the analgesic peptide gene BmK AngM1 from Buthus martensii Karsch
Abstract:

Codon bias is an important factor which influences heterologous gene expression.  Optimizing codon sequence could improve expression level of heterologous gene.  In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris.  The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one.  These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.

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