| 木质素过氧化物酶基因（LipH8）的克隆及在甲醇毕赤酵母中的表达 |
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| 引用本文： | 王海宽,孙亚范,杜连祥,路福平.木质素过氧化物酶基因（LipH8）的克隆及在甲醇毕赤酵母中的表达[J].微生物学报,2004,44(2):258-260. |
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| 作者姓名： | 王海宽 孙亚范 杜连祥 路福平 |
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| 作者单位： | 天津科技大学食品科学与生物工程学院,天津,300222 |
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| 摘 要： | 以黄孢原毛平革菌 (Phanerochaetechrysosporium)RNA为模板 ,克隆LipH8基因片段 ,研究LipH8基因在甲醇毕赤酵母中的表达。构建了甲醇酵母表达质粒pMETA_LipH8载体 ,并将其线性化后用电穿孔法导入PichiamethabolicaPMAD16 ,部分阳性克隆的PCR结果表明LipH8基因已经整合到甲醇毕赤酵母染色体上 ,经摇瓶培养筛选出表达水平较高的酵母工程菌株。胞外木质素过氧化物酶活力达 932U L。
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| 关 键 词： | cDNA 异源表达 木质素过氧化物酶 甲醇毕赤酵母 |
| 文章编号： | 0001-6209(2004)02-0258-03 |
| 修稿时间： | 2003年6月10日 |
Cloning of cDNA Encoding Lignin Peroxidase (LipH8) from Phanerochaete chrysosporium and Expression in Pichia methanolica |
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| WAHG Hai_Kuan,SUN Ya_Fan,DU Lian_Xiang,LU Fu_Ping.Cloning of cDNA Encoding Lignin Peroxidase (LipH8) from Phanerochaete chrysosporium and Expression in Pichia methanolica[J].Acta Microbiologica Sinica,2004,44(2):258-260. |
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| Authors: | WAHG Hai_Kuan SUN Ya_Fan DU Lian_Xiang LU Fu_Ping |
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| Institution: | WAHG Hai_Kuan SUN Ya_Fan DU Lian_Xiang LU Fu_Ping * |
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| Abstract: | The expression of lignin peroxidase gene in Pichia methanolica was investigated. The cDNA encoding lignin peroxidase was cloned by RT_PCR using RNA of P.chrysosporium as template. The lignin peroxidase cDNA was inserted into plasmid pMETA and transformed into Pichia methanolica pMAD16. PCR results indicated that the lignin peroxidase gene from P.chrysosporium has been successfully cloned into Pichia methanolica. The transforments was screened to get high expression strain by shake_flask culture. The extracellular lignin peroxidase activity in this primary construct was 932U/L. |
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| Keywords: | cDNA cloning Heterologous expression Lignin peroxidase Pichia methanolica |
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