杂色云芝漆酶基因(Lcc1)的克隆及在甲醇毕赤酵母中的表达  

CLONING OF cDNA ENCODING LACCASE (Lccl) FROM CORIOLUS VERSICOLOR AND EXPRESSION OF THE GENE IN PICHIA METHANOLICA

作　　者：郭梅[1,2] 蒲军[1] 杜连祥[1] 路福平[1] 白东清[1,3] 

机构地区：[1]天津科技大学生物工程学院工业微生物重点实验室 [2]天津农学院食品科学系天津300384 [3]天津农学院水产系天津300384

出　　处：《菌物学报》2005年第2期221-226,共6页Mycosystema

摘　　要：以白腐菌杂色云芝Coriolus versicolor RNA为模板,通过RT-PCR获得漆酶Leel基因的cDNA片段。构建了甲醇酵母表达质粒pMETA-Lccl载体,并将其线性化后用电穿孔法导入Pichia methabolica PMAD16,部分阳性克隆的PCR结果表明Lccl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。The expression of laccase gene in Pichia methanolica was investigated. The cDNA encoding laccase was cloned by RT-PCR using RNA of the white rot fungus Coriolus versicolor as template. The lassase cDNA was inserted into plasmid pMETA and transformed into Pichia methanolica PMAD16. PCR results indicated that the lignin peroxidase gene from Coriolus versicolor has been successfully cloned into Pichia methanolica. The transforments was screened to get high expression strain by shake-flask culture. The extracellular laccase activity in this primary construct was 53U/L.

关 键 词：甲醇毕赤酵母 杂色云芝 酶基因 cDNA片段 RT-PCR PICHIA E1基因 表达质粒 甲醇酵母 电穿孔法 阳性克隆 工程菌株 摇瓶培养 白腐菌 RNA 线性化 染色体 酶活力 漆酶 载体 

分 类 号：Q943.2[生物学—植物学]