球孢白僵菌降解昆虫体壁蛋白酶基因 CDEP-1的克隆与序列分析 |
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引用本文: | 方卫国,张永军,杨星勇,王中康,裴炎. 球孢白僵菌降解昆虫体壁蛋白酶基因 CDEP-1的克隆与序列分析[J]. 遗传学报, 2002, 29(3): 278-282 |
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作者姓名: | 方卫国 张永军 杨星勇 王中康 裴炎 |
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作者单位: | 西南农业大学生物技术中心,重庆北碚,400716 |
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基金项目: | 国家自然科学基金资助项目 (项目编号 :3 9880 0 1),国家高技术研究发展计划 (863 )资助(批准号 :2 0 0 1AA2 14 0 5 1)~~ |
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摘 要: | 构建了球孢白僵菌不同诱导时间的混合cDNA文库。根据丝氨酸类蛋白酶的保守区域设计引物,以构建cDNA文库同样的mRNA为模板,采用RT-PCR法得到长度为594bp的片段BbP。序列测定表明BbP是球孢白僵菌类枯草杆菌蛋白酶Prl的一部分,以BbP为探针,从上述cDNA文库筛选得到长度为1557bp的克隆CDEP-1。CDEP-1含有一个1134bp的开放阅读框(ORF),编码377个氨基酸,分子量为38616、PI=8.302的蛋白酶前体。CEDP-1的核苷酸序列与蛋白酶K、金龟子绿僵菌Prl、球孢白僵菌Prl的同源性分别为:57.9%、54.7%、83.3%。根据cDNA序列扩增到CDEP-1的基因组序列,分析表明其中含有3个内含子。Southern杂交表明CDEP-1在球孢白僵菌是单拷贝。
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关 键 词: | 球孢白僵菌 类枯草杆菌蛋白酶 CDEP-1 昆虫体壁 侵染 降解 基因克隆 序列分析 |
文章编号: | 0379-4172(2002)03-0278-05 |
修稿时间: | 2001-09-17 |
Cloning and CharacteriZation of Cuticle Degrading Enzyme CDEP-1 from Beauveria bassiana |
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Abstract: | Beauveria bassiana extracellular subtilisin like serine endoprotease is a potential virulence factor by virtue of its activity against insect cuticles. A cDNA library was constructed using mRNA from mycelia of Beauveria bassiana grown on cuticle/chitin cultures. A cDNA clone of the protease, designated CDEP 1, was isolated from cDNA library. CDEP 1 contained an 1?134bp ORF that predicted a protein of 377 amino acids with M r=38?616 and PI=8.302. The amino acid sequence of the gene shows 57.9%,83.3% and 54.7% identity to Metarhizium nisopliae Pr1, Beauveria bassiana Pr1 and proteinase K, respectively. Southern analysis indicated that CDEP 1 was present as singly copy in Beauveria bassiana. |
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Keywords: | Beauveria bassiana subtilisin like serine endoprotease CDEP 1 |
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