| 灰盖拟鬼伞核定位蛋白重组表达系统的构建 |
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| 引用本文： | 王宇,赵静,田宣,金盛宇,冉妍妍,黄小娟,袁静,袁生,刘中华.灰盖拟鬼伞核定位蛋白重组表达系统的构建[J].菌物学报,2022,41(1):68-77. |
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| 作者姓名： | 王宇 赵静 田宣 金盛宇 冉妍妍 黄小娟 袁静 袁生 刘中华 |
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| 作者单位： | 南京师范大学 生命科学学院 江苏省微生物与功能基因组学重点实验室 江苏省微生物资源产业化工程技术研究中心,江苏 南京 210023 |
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| 基金项目： | 国家自然科学基金(31600053)。 |
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| 摘 要： | 为构建灰盖拟鬼伞Coprinopsis cinerea的核定位蛋白重组表达系统,本研究通过蛋白序列比对和信息学分析,预测了灰盖拟鬼伞组蛋白H2B的核定位序列,构建了融合组蛋白H2B核定位序列的绿色荧光蛋白(green fluorescent protein,GFP)重组表达载体,将该载体转入灰盖拟鬼伞AmutBmut菌...
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| 关 键 词： | 核定位序列 重组表达 启动子 绿色荧光蛋白 |
| 收稿时间： | 2021-05-14 |
Construction of the recombinant nuclear localization expression system in Coprinopsis cinerea |
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| Authors: | WANG Yu ZHAO Jing TIAN Xuan JIN Shengyu RAN Yanyan HUANG Xiaojuan YUAN Jing YUAN Sheng LIU Zhonghua |
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| Institution: | Jiangsu Key Laboratory for Microbes and Microbial Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, College of Life Sciences, Nanjing Normal University, Nanjing 210023, Jiangsu, China |
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| Abstract: | To construct a recombinant nuclear localization (NL) expression system for Coprinopsis cinerea, the NL sequence (NLS) of histone H2B was determined by amino acid sequence alignment and informatics analysis. The recombinant expression vector of fusion green fluorescent protein (GFP) with the NLS of histone H2B was constructed and transferred into C. cinerea AmutBmut strain to examine whether this NLS can guide GFP expression to the nucleus. The results showed that the codon-optimized GFP gene was expressed to produce green fluorescence under the control of the β-tubulin promoter of C. cinerea. Furthermore, NLS from histone H2Bb (CC1G_07639) can successfully induce GFP expression in the nucleus. In this study, a recombinant NL expression system for C. cinerea with proven efficiency was constructed. Our findings are beneficial to optimizing the CRISPR/Cas9 gene editing system and further exploring the mechanisms of fruiting body formation in C. cinerea. |
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| Keywords: | nuclear localization sequence recombinant expression promoter green fluorescent protein |
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