交链孢一种真菌激活蛋白的分离纯化及其基因的克隆与表达-环境科学专业论文.docxVIP

摘 摘 要 本论文介绍了生物农药的定义及分类。对国内外新型微生物蛋白农药的研究进展 进行了综述，真菌激活蛋白属于一种新型、广谱、高效、多功能、无残留、对天敌安 全及与环境相容生物农药。其作用机理在性质上类似动物免疫的一种抗病机制。对微 生物蛋白农药的发展趋势和应用前景进行了概述。 从一种真菌中初步分离纯化了分子量约为42KD的激活蛋白，并对其氨基酸序列 进行了测序，运用PCR技术获得了此激活蛋白的部分编码基因np45及其编码的氨基 酸为NP45，构建了融合表达载体，并在大肠杆菌中进行了成功的表达。 np45及NP45在Gene Bank中比对后发现，NP45只与根瘤菌属Bradyrhizobium japonicum USDA 110编码的序列号为blr0937的蛋白具有19．14％的同源性，而与 Aspergillus nidulans FGSC A4编码的蛋白，序列号为AN0215．2的同源性为53．78％。 NP45与过敏蛋I!tHarpin、隐地蛋I；tElicitin无同源性。此基因属于新基因，此蛋白属于 新蛋白。 关键词：微生物蛋白农药；过敏蛋白；隐地蛋白；激活蛋白；基因克隆；融合表达 V AbstractIn Abstract In this paper,the definition and classification of biopesticide was introduced，and the research development ofnew microbial proteinaceous pesticide reviewed．Fungal activator is a novel microbial proteinaceous pesticide，which is multifunctional，high efficient，no residue，environment-friendly,and safety to natural enemy．Its action mechanism is similar to that of animal immunity．In this article，the developmental trend and application potential of this new microbial protein pesticide was reviewed． In this study，a fungal activator,approximately Mr．42KD，was purified from Alternaria axo strain and sequenced．A partial fungal activator gene np45 was obtained by using PCR technique．Bacterial fusion expressed vectors of np45 gene was constructed and successfully expressed in E．coli，named by NP45．Analysis of NP45 at GeneBank Blast showed that the identity between NP45 and USDA 110 blr0937 from Bradyrhizobium japonieum was 19．14％，the highest score in bacteria．Furthermore，the identity of NP45 and FGSC A4 sequence number AN0215．2 from Aspergillus nidulans was 53．78％，the highest score in fungi．There was no homology between NP45 and other known microbial proteinaceous pesticides，such as hairpin，elicitin．This result indicated that，rip45 was a new kind of gene，and NP45 a novel protein． Keywords：microbial protein pesticide；harpin；elicitin；activator；gene cloning；fusion expression V 独创性声明本人声}抛所呈交的论文是我个人在导哪指导F进仃的研究工作及取得的研究 独创性声明 本人声}抛所呈交的论文是我个人在导哪指导F进仃的研究工作及取得的研究 成小。N我