摘要
PCR扩增得到微小毛霉凝乳酶结构基因并测定其核苷酸序列,用DNAman软件分析其核苷酸序列及推衍得到的多肽序列,结果表明扩增得到的片段为凝乳酶基因。构建了重组菌Pichia pastoris KM71/PIC9K-mcp,通过G418抗性筛选得到具有多拷贝基因的整合重组菌MK3。用甲醇诱导进行初步发酵试验,测得重组菌MK3酶活为5.4U/ml。
In this paper, rennin gene was amplified from Mucor pusillus and sequenced. The nucleic acid sequence and amino-acid sequence encoded by the gene were compared using DNAman biological software. Results showed that amplified fragmentwas a novel rennin gene(mcp). re-combination Pichia pastoris KM71/PIC9K-mcp was constructed successfully and obtained amulti-copied integrated strain of MK3 by resistant screening (G418). Initial fermation experiments were carried out andmethanol was used as sole carbon source. The enzyme activity of re-combination MK3 was 5.4U/ml in above condition.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2005年第7期58-63,共6页
Food Science
关键词
凝乳酶
克隆
序列分析
重组菌
表达
rennin
clone
sequence analysis
re-combination strain
express
