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凝乳酶的基因克隆、序列分析及初步表达

Molecular Cloning, Sequence Analysis and Primary Expression of Mucor pusillus DNA

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【作者】 邱重晏徐敏王正祥

【Author】 QIU Chong-yan,XU Min,WANG Zheng-xiang(The Key Laboratory of Industrial Biology, School of Biotechnology, Southern Yangtze University,Wuxi 214036, China)

【机构】 江南大学生物工程学院工业生物技术教育部重点实验室江南大学生物工程学院工业生物技术教育部重点实验室 江苏无锡214036江苏无锡214036江苏无锡214036

【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。

【Abstract】 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.

【关键词】 凝乳酶克隆序列分析重组菌表达
【Key words】 renninclonesequence analysisre-combination strainexpress
  • 【分类号】Q785
  • 【被引频次】24
  • 【下载频次】352
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