| 黑曲霉外切葡聚糖酶基因密码子的优化及表达 |
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| 引用本文： | 李国庆,范松,裴建军,赵林果,高树峰. 黑曲霉外切葡聚糖酶基因密码子的优化及表达[J]. 南京林业大学学报(自然科学版), 2012, 36(4): 84-88 |
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| 作者姓名： | 李国庆 范松 裴建军 赵林果 高树峰 |
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| 作者单位： | 南京林业大学化学工程学院,江苏省生物质绿色燃料与化学品重点实验室,江苏南京210037 |
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| 基金项目： | 国家自然科学基金项目，江苏高校优势学科建设工程资助项目 |
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| 摘 要： | 通过NCBI(美国国立生物技术信息中心)对黑曲霉的来源外切葡聚糖酶基因(cbh1)的氨基酸序列进行分析,发现其16个活性部位中有6个位点附近的氨基酸为毕赤酵母(Pichia pasto-ris)表达的稀有密码子。利用反向长距离PCR技术将这些活性位点附近的稀有密码子定点突变为毕赤酵母表达偏爱密码子。将优化后的表达载体pPICZαA-cbh1m转入毕赤酵母KM71H菌株中进行诱导表达。经密码子优化后的cbh1m基因较未优化前cbh1基因的表达量提高了17%。
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| 关 键 词： | 外切葡聚糖酶 定点突变 密码子优化 黑曲霉 毕赤酵母 |
Codon optimization of cellobiohydrolase gene of Aspregillus niger and its expression in Pichia pastoris |
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| LI Guoqing , FAN Song , PEI Jianjun , ZHAO Linguo , GAO Shufeng. Codon optimization of cellobiohydrolase gene of Aspregillus niger and its expression in Pichia pastoris[J]. Journal of Nanjing Forestry University(Natural Sciences ), 2012, 36(4): 84-88 |
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| Authors: | LI Guoqing FAN Song PEI Jianjun ZHAO Linguo GAO Shufeng |
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| Affiliation: | (College of Chemical Engineering,Nanjing Forestry University,Jiangsu Key Lab of Biomass-based Green Fuels and Chemicals,Nanjing 210037,China) |
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| Abstract: | NCBI analysis revealed that there were sixteen active sites in cellobiohydrolase gene(cbh1) from Aspregillus niger,and were found rare codons around six active sites.To improve expression efficiency of recombinant cellobiohydrolase in Pichia pastoris KM71H,those rare codons around active site of cbh1 were replaced with preferable codons used in P.pastoris by long-distance inverse PCR.The recombinant plasmid pPICZαA-cbh1m which contained the modified cbh1 sequence was transformed in P.pastoris KM71H.After methanol induction,the expression level of cbh1m was 17 % higher than that of cbh1. |
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| Keywords: | cellobiohydrolase site-directed mutagenesis codon optimization Aspregillus niger Pichia pastoris |
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