| 里氏木霉RutC30常温常压等离子体（ARTP）诱变筛选pyr4基因缺陷型菌株及转化系统的建立 |
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| 引用本文： | 安莉颖,易欣欣,秦丽娜,施思,陶勇,伍红,董志扬. 里氏木霉RutC30常温常压等离子体（ARTP）诱变筛选pyr4基因缺陷型菌株及转化系统的建立[J]. 菌物学报, 2014, 33(6): 1272-1280. DOI: 10.13346/j.mycosystema.130071 |
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| 作者姓名： | 安莉颖 易欣欣 秦丽娜 施思 陶勇 伍红 董志扬 |
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| 作者单位： | 1西南民族大学生命科学与技术学院 四川 成都 610041;2中国科学院微生物研究所 北京 100101;3北京农学院食品科学与工程学院 北京 102206 |
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| 基金项目： | 国家自然科学基金(No.30970073);国家重点基础研究发展规划(973项目)(No.2011CB707402);国家高技术研究发展计划(863计划)(No.2012AA092103) |
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| 摘 要： | 以里氏木霉Trichoderma reesei重要工业生产菌RutC30为出发菌株，通过等离子体（ARTP）诱变筛选，以5-氟乳清酸（5-FOA）和尿苷（Uridine）进行筛选，获得一株pyr4基因缺陷株RutC30ΔU3。用含有野生型里氏木霉pyr4基因的互补质粒转化突变株，可回复野生性状。经测序发现其pyr4基因在核酸序列多个位点发生突变，其中包括两个错义突变和一个移码突变，从而导致乳清酸核苷-5′-磷酸脱羧酶失活。经遗传稳定性研究分析，传代5次后仍保持良好的尿苷依赖性、去葡萄糖阻遏以及高产纤维素酶特性。经实验筛选获得了pyr4基因缺陷菌株可作为基因表达系统的受体菌株，建立了以尿苷营养缺陷为筛选标记的木霉转化系统。
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| 关 键 词： | 里氏木霉 等离子体 诱变 pyr4基因 |
Isolation of Trichoderma reesei RutC30 pyr4 gene mutant by atmos-pheric room temperature plasma (ARTP) and establishment of a trans-formation system |
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| AN Li-Ying,YI Xin-Xin,QIN Li-Na,SHI Si,TAO Yong,WU Hong,DONG Zhi-Yang. Isolation of Trichoderma reesei RutC30 pyr4 gene mutant by atmos-pheric room temperature plasma (ARTP) and establishment of a trans-formation system[J]. Mycosystema, 2014, 33(6): 1272-1280. DOI: 10.13346/j.mycosystema.130071 |
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| Authors: | AN Li-Ying YI Xin-Xin QIN Li-Na SHI Si TAO Yong WU Hong DONG Zhi-Yang |
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| Affiliation: | 1College of Life Science and Technology, Southwest University for Nationalities, Chengdu, Sichuan 610041, China;2Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;3Food Science and Engineering College, Beijing University of Agriculture, Beijing 102206, China |
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| Abstract: | An uridine auxotrophic mutant of Trichoderma reesei RutC30 was isolated by resistance to 5-fluoroorotic acid through Atmospheric Room Temperature Plasma mutagenesis. The mutant, called RutC30ΔU3, recovered to uridine prototrophy by transformation with the wild type T. reesei pyr4 gene cassette. Sequence analysis of this mutated gene indicated that two missenses mutation and one frame-shift mutation had occurred. Mitotic stability analysis of the mutant strain RutC30ΔU3 showed a quite good genetic stability. In this study, the pyr4 mutant RutC30ΔU3 was successfully established and proved to be an excellent recipient strain for gene expression. |
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| Keywords: | Trichoderma reesei atmospheric room temperature plasma mutation pyr4 gene |
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