漆酶基因的克隆及在甲醇毕赤酵母中的表达  

MOLECULAR CLONING OF THE CDNA ENCODING LACCASE AND EXPRESSION IN PICHIA METHANOLICA

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

机构地区：[1]天津农学院食品科学系,天津300384 [2]天津科技大学生物工程学院工业微生物重点实验室,天津300222 [3]天津农学院水产系,天津300384

出　　处：《食品研究与开发》2008年第12期30-32,共3页Food Research and Development

基　　金：天津市自然科学基金重点项目(07JCZDJC03100)

摘　　要：以白腐菌杂色云芝RNA为模板,通过RT-PCR获得不带自身信号肽漆酶Lcc1基因的cDNA片段。构建了重组表达载体pMETαA-Lcc1,并将其线性化后用电穿孔法导入Pichia methabolica PMAD16,PCR结果表明Lcc1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菌株,漆酶酶活力达78U/L。The cDNA encoding laccase without the native secretion was cloned by RT-PCR using RNA of the white rot fungus Coriolus versicolor as template. The lassase cDNA was inserted into plasmid pMETαA and transformed into Pichia methanolica PMAD16. PCR results indicated that the Lccl gene from C. versicolor has been successfully cloned into P. methanolica. The transforments was screened to get high expression strain by shake-flask culture. The extracellular laccase activity in this primary construct was 78 U/L.

关 键 词：漆酶 CDNA 甲醇毕赤酵母 异源表达 

分 类 号：TS201.25[轻工技术与工程—食品科学]