粗糙脉孢霉酸性蛋白酶基因的克隆与鉴定  

Cloning and Identification of a Gene Encoding an Acidic Protease from Neurospora crassa

作　　者：邱重晏[1] 王正祥[1] 

机构地区：[1]工业生物技术教育部重点实验室和江南大学生物工程学院

出　　处：《微生物学通报》2006年第4期91-94,共4页Microbiology China

基　　金：新世纪优秀人才支持计划(No.NCET04-9704)

摘　　要：以粗糙脉胞霉CICIM F00021染色体DNA为模板,通过PCR扩增得到粗糙脉胞霉酸性蛋白酶结构基因。对其序列进行了测定和分析,表明扩增得到的片段为酸性蛋白酶基因。将扩增出的酸性蛋白酶基因克隆入酵母表达载体中获得重组质粒pPIC9K-ap。将其转化入毕赤氏酵母获得重组菌NA3。重组酸性蛋白酶在pH4·0和45oC下表现出最高活性。A gene encoding a putative acidic protease was isolated from Neurospora crassa chromosomal DNA. The nucleic acid sequencing and analysis indicated the amplified fragment was an acidic protease gene （ap）. Purified PCR product was subcloned into a yeast expression vector, yielding recombinant pPICgK-ap. The recombinant NA3 was constructed by transforming pPIC9K-ap into Pichia pastoris KM71. The recombinant acidic protease produced by P. pastoris performed the highest activity at pH 4. 0 and 45℃.

关 键 词：粗糙脉胞霉 酸性蛋白酶 克隆 序列分析 表达 

分 类 号：Q78[生物学—分子生物学]