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Position: Home > Articles > Molecular Cloning, Sequence Analysis and Primary Expression of Mucor pusillus DNA FOOD SCIENCE 2005,26 (7) 58-63

凝乳酶的基因克隆、序列分析及初步表达

作  者:
邱重晏;徐敏;王正祥
单  位:
江南大学生物工程学院工业生物技术教育部重点实验室
关键词:
凝乳酶;克隆;序列分析;重组菌;表达
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。
译  名:
Molecular Cloning, Sequence Analysis and Primary Expression of Mucor pusillus DNA
作  者:
QIU Chong-yan,XU Min,WANG Zheng-xiang(The Key Laboratory of Industrial Biology, School of Biotechnology, Southern Yangtze University,Wuxi 214036, China)
关键词:
rennin;clone;sequence analysis;re-combination strain;express
摘  要:
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.

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