| 少孢节丛孢菌Aoz1基因的生物信息学分析及原核表达条件的优化 |
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| 引用本文： | 尹相吉,王新芳,聂福贵,郭少平,钱英红,陈湘江,乔一丹,李泽浩,符江涛,杨晓野,王瑞.少孢节丛孢菌Aoz1基因的生物信息学分析及原核表达条件的优化[J].中国兽医科学,2019(3):333-340. |
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| 作者姓名： | 尹相吉 王新芳 聂福贵 郭少平 钱英红 陈湘江 乔一丹 李泽浩 符江涛 杨晓野 王瑞 |
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| 作者单位： | 内蒙古农业大学兽医学院农业部动物疾病临床诊疗技术重点实验室;内蒙古阿拉善盟畜牧兽医工作站;内蒙古自治区农牧业科学院;山东省诸城市畜牧兽医管理局 |
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| 基金项目： | 国家重点研发计划项目(2017YFD0501200);国家自然科学基金资助项目(31460656);内蒙古自然科学基金资助项目(2014MS0339);家畜疫病病原生物学国家重点实验室开放基金项目(SKLVEB2017KFKT010) |
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| 摘 要： | 为获悉少孢节丛孢菌内蒙古株Aoz1基因的详细信息,以研究捕食线虫性真菌对线虫的作用机制,本试验以捕食线虫性真菌的代表种——少孢节丛孢菌内蒙古株为研究对象,首先扩增其胞外丝氨酸蛋白酶的基因编码序列,并进行生物信息学分析,然后将其构建至原核表达载体pET-32a中,再转化至Transetta(DE3)表达感受态细胞中,进行表达条件的优化以及表达产物的纯化。结果表明,少孢节丛孢菌内蒙古株丝氨酸蛋白酶Aoz1基因与已发表的丝氨酸蛋白酶Aoz1基因的同源性为99%,其蛋白具备螺旋结构;在原核系统中进行表达,其最适IPTG诱导浓度为1.5 mmol/L,最佳诱导时间为8 h,确定了重组蛋白的最佳表达条件,并获得了重组蛋白。对该酶基因的分析,为后续有关蛋白酶和捕食线虫性真菌杀灭寄生性线虫机理方面的研究提供了参考资料。
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| 关 键 词： | 捕食线虫性真菌 少孢节丛孢菌 丝氨酸蛋白酶 Aoz1基因 |
Bioinformatics analysis of Aoz1 gene of Arthrobotrys oligospora and optimization of its prokaryotic expression conditions |
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| YIN Xiang-ji,WANG Xin-fang,NIE Fu-gui,GUO Shao-ping,QIAN Ying-hong,CHEN Xiang-jiang,QIAO Yi-dan,LI Ze-hao,FU Jiang-tao,YANG Xiao-ye,WANG Rui.Bioinformatics analysis of Aoz1 gene of Arthrobotrys oligospora and optimization of its prokaryotic expression conditions[J].Veterinary Science in China,2019(3):333-340. |
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| Authors: | YIN Xiang-ji WANG Xin-fang NIE Fu-gui GUO Shao-ping QIAN Ying-hong CHEN Xiang-jiang QIAO Yi-dan LI Ze-hao FU Jiang-tao YANG Xiao-ye WANG Rui |
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| Institution: | (Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease,the Ministry of Agriculture,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;The Animal Husbandry and Veterinary WorkingStation of Alashan League of Inner Mongolia,Alashim Zuo Banner 750306,China;Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China;Zhucheng Animal Husbandry and Veterinary Administration Bureau of Shandong Province,Weifang 262200,China) |
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| Abstract: | YIN Xiang-ji;WANG Xin-fang;NIE Fu-gui;GUO Shao-ping;QIAN Ying-hong;CHEN Xiang-jiang;QIAO Yi-dan;LI Ze-hao;FU Jiang-tao;YANG Xiao-ye;WANG Rui(Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease,the Ministry of Agriculture,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;The Animal Husbandry and Veterinary WorkingStation of Alashan League of Inner Mongolia,Alashim Zuo Banner 750306,China;Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences,Hohhot 010031,China;Zhucheng Animal Husbandry and Veterinary Administration Bureau of Shandong Province,Weifang 262200,China) |
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| Keywords: | nematode trapping fungi Arthrobotrys oligospora serine protease Aoz1 gene |
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