| 人溶菌酶基因在毕赤酵母中的表达及其抗菌活性检测 |
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| 引用本文： | 夏清风,侯英敏,曹芳,金朝霞.人溶菌酶基因在毕赤酵母中的表达及其抗菌活性检测[J].大连轻工业学院学报,2011(5). |
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| 作者姓名： | 夏清风 侯英敏 曹芳 金朝霞 |
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| 作者单位： | 大连工业大学生物工程学院; |
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| 基金项目： | 大连市优秀青年人才基金项目(2009J22DW036) |
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| 摘 要： | 将PCR扩增的人溶菌酶编码基因克隆到毕赤酵母分泌型表达载体pPIC9K中,双酶切和测序鉴定表明,重组载体构建正确。电转化毕赤酵母GS115菌株,通过G418筛选获得高拷贝重组菌株后,用甲醇诱导表达;通过SDS-PAGE检测证明上清中有目的蛋白,表达产物与预期大小的人溶菌酶蛋白分子质量15ku一致,其表达量占分泌总蛋白的32%。体外抑菌实验证实重组人溶菌酶蛋白对溶壁微球菌、金黄色葡萄球菌、大肠杆菌和枯草杆菌均有明显的抑菌作用,表明在毕赤酵母中成功表达了重组人溶菌酶。
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| 关 键 词： | 人溶菌酶 毕赤酵母 pPIC9K 表达 |
Expression of the human lysozyme in Pichia pastoris and antibacterial activities test |
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| XIA Qing-feng,HOU Ying-min,CAO Fang,JIN Zhao-xia School of Bioengineering,Dalian Polytechnic University,Dalian ,China.Expression of the human lysozyme in Pichia pastoris and antibacterial activities test[J].Journal of Dalian Institute of Light Industry,2011(5). |
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| Authors: | XIA Qing-feng HOU Ying-min CAO Fang JIN Zhao-xia School of Bioengineering Dalian Polytechnic University Dalian China |
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| Affiliation: | XIA Qing-feng,HOU Ying-min,CAO Fang,JIN Zhao-xia School of Bioengineering,Dalian Polytechnic University,Dalian 116034,China |
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| Abstract: | The HLY coding gene was cloned into P.pastoris expression vector pPIC9K.The pPIC9K/HLY was verified by two enzyme digestion and sequencing and then transferred into P.pastoris GS115 strain by electroporation.The recombinant strains were isolated by G418.The results of SDS-PAGE demonstrated that the fermentation supernatant contained the recombinant protein up to 32% of total proteins and its molecular weight is 15 ku.In vitro the recombinant protein has certain antimicrobial activity against M.lysodeikticus... |
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| Keywords: | human lysozyme Pichia pastoris pPIC9K expression |
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