| 何晓红1，杨宏国1，伍晓丽2，刘 飞2，谢永芳1，蒋龙星1，王 宇1，冉玲芳1.虫草棒束孢类枯草杆菌蛋白酶基因克隆及分析[J].广东农业科学,2017,44(5):32-36 |
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虫草棒束孢类枯草杆菌蛋白酶基因克隆及分析 |
| Cloning and analysis of encoding cDNA sequence of sublitisin-like protease gene from Isaria farinosa |
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| DOI：10.16768/j.issn.1004-874X.2017.05.006 |
| 中文关键词: 虫草棒束孢 虫生真菌 蛋白酶 cDNA |
| 英文关键词: Isaria farinosa entomogenous fungi subtilisin-like protease complementary DNA |
| 基金项目:国家自然科学基金（30901964，81072997，81173479）；重庆市基础与前沿研究项目（cstc2015J
A80006）；重庆市基本科研业务费计划项目（2015cstc-jbky-01912）；重庆市教委科学技术研究项目（KJ1500420，
KJ130520）；重庆市大学生创新创业训练计划项目（201610617103） |
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| 中文摘要: |
| 采用逆转录PCR 方法克隆获得1 个虫草棒束孢类枯草杆菌蛋白酶基因的开放阅读框（ORF）
序列，并对其进行生物信息学分析。该基因的开放阅读框全长1 599 bp，编码的蛋白质由532 个氨基酸组成
（包括18 个氨基酸的信号肽）。预测蛋白质的等电点和分子量大小分别为6.256、56.936 ku。进行序列分析
和进化树构建，发现该蛋白酶与已知的几个种类枯草杆菌蛋白酶具有很高的同源性，且与虎甲寄生菌的粉
拟青霉位于同一进化分支。实验首次鉴定了虫草棒束孢类枯草杆菌蛋白酶基因的编码区序列，为进一步研
究虫草棒束孢致死蝙蝠蛾幼虫的作用机理提供理论依据。 |
| 英文摘要: |
| In the present study，the coding cDNA sequence of Isaria farinosa subtilisin-like protease（IsSUB）
was cloned by reverse transcription PCR and analyzed by bioinformatical tools in detail. The subtilisin contains 1 599
nucleotides with a putative open reading frame（ ORF） encoding 532 amino acid residues（ with a signal peptide
of 18 amino acid residues）. The isoelectric point（ pI） and predicted molecular weight of IsSUB were 6.256 and
56.936 kDa，respectively. Amino acid sequence and phylogenetic tree analysis among I. farinosa and other insects
represented that these fungal subtilisin had a high homology，and IsSUB existed in one evolutionary branching with
Cicindelidae sp. Paecilomyces farinosus. It was concluded that the ORF of subtilisin-like protease from I. farinosa
provided theoretical basis for damaging on Hepialus larvae body wall. |
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