去信号肽的木质素过氧化物酶基因在甲醇毕赤酵母中的表达  

Expression of lignin peroxidase of Phanerochaete chrysosporium without signal peptide in Pichia methanolica

作　　者：王海宽[1] 路福平[1] 孙亚范 杜连祥[1] 

机构地区：[1]天津科技大学生物工程学院,天津300222

出　　处：《云南大学学报（自然科学版）》2004年第S2期87-90,共4页Journal of Yunnan University(Natural Sciences Edition)

摘　　要：研究了LipH8基因在甲醇毕赤酵母中的表达.以黄孢原毛平革菌(Phanerochaetechrysosporium)RNA为模板,克隆LipH8基因片段.构建了甲醇酵母表达质粒pMETαA-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1933U/L.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 pMETα A 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 1 933 U/L.

关 键 词：CDNA 异源表达 木质素过氧化物酶 甲醇毕赤酵母 

分 类 号：Q786[生物学—分子生物学]