| 牛凝乳酶基因在毕赤酵母中的重组表达 |
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| 引用本文： | 张莉,姜媛媛,张健,杨贞耐.牛凝乳酶基因在毕赤酵母中的重组表达[J].生物工程学报,2009,25(8):1160-1165. |
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| 作者姓名： | 张莉 姜媛媛 张健 杨贞耐 |
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| 作者单位： | 中国农业科技东北创新中心农产品加工研究中心,长春,130033 |
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| 基金项目： | 国家高技术研究发展计划(863计划)(No.2006AA10Z306);;吉林省科技发展计划项目(Nos.20060219,20080228);;现代农业产业技术体系建设专项资金(农科教发[2007]14号)资助~~ |
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| 摘 要： | 通过PCR技术从克隆载体pMD18T-Prochy上扩增牛凝乳酶原基因,双酶切后定向插入到酵母表达载体pPICZaA中,构建表达质粒pPICZaA-Prochy,线性化后电转化毕赤酵母GS115,经PCR和测序鉴定凝乳酶原基因成功插入到毕赤酵母的基因组中。在甲醇诱导下进行凝乳酶的表达,SDS-PAGE分析证明重组凝乳酶的分子量约为37 kD,培养基上清液中凝乳酶的活性为12.2 SU/mL。本研究首次应用毕赤酵母表达牛凝乳酶,在培养基中获得分泌表达的重组凝乳酶,为干酪工业提供了新型及优良的凝乳酶来源。
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| 关 键 词： | 牛凝乳酶 分泌表达 毕赤酵母 |
| 收稿时间： | 2009/4/20 0:00:00 |
Recombinant expression of bovine chymosin in Pichia pastoris |
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| Li Zhang,Yuanyuan Jiang,Jian Zhang and Zhennai Yang.Recombinant expression of bovine chymosin in Pichia pastoris[J].Chinese Journal of Biotechnology,2009,25(8):1160-1165. |
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| Authors: | Li Zhang Yuanyuan Jiang Jian Zhang and Zhennai Yang |
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| Institution: | Center of Agro-food Technology, Northeast Agricultural Research Center of China, Changchun 130033, China;Center of Agro-food Technology, Northeast Agricultural Research Center of China, Changchun 130033, China;Center of Agro-food Technology, Northeast Agricultural Research Center of China, Changchun 130033, China;Center of Agro-food Technology, Northeast Agricultural Research Center of China, Changchun 130033, China |
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| Abstract: | To express bovine chymosin in yeast,we amplified the prochymosin gene from the plasmid pMD18T-Prochy by PCR,and then cloned the gene into the expression vector pPICZaA,resulting in pPICZaA-Prochy.Pichia pastoris GS115 was used as host cells.Integration of the prochymosin cDNA into the Pichia pastoris genome was confirmed by PCR and sequencing analysis.Chymosin was expressed in Pichia pastoris successfully,and a strong band at about 37 kD was shown by SDS-PAGE.Activity tests showed that the chymosin activity... |
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| Keywords: | bovine chymosin secretory expression Pichia pastoris |
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