| 沈慧敏, 李 超, 高 利*, 刘太国, 刘 博, 陈万权.原生质体法介导真菌遗传转化的研究进展[J].植物保护,2017,43(2):25-28. |
| 原生质体法介导真菌遗传转化的研究进展 |
| Research progress in transformation of fungi mediated by protoplasts |
| 投稿时间：2016-03-23 修订日期：2016-07-11 |
| DOI： |
| 中文关键词: 原生质体 制备与再生 PEG介导 电击 REMI 转化子 |
| 英文关键词:protoplast preparation and regeneration PEG-mediated electrical stimulation REMI transformant |
| 基金项目:国家自然科学基金(31571965)；北京市自然科学基金(6162022)；北京市科技新星(Z131105000413057)；北京市自然科学基金北京市科学技术研究院联合资助项目(L140012)；小麦产业技术体系(CARS-03) |
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| 中文摘要: |
| 原生质体法介导真菌遗传转化体系是实现大规模随机、定点突变和菌种改良的方法之一, 为研究真菌致病性和侵染机制奠定了基础。原生质体法构建转化体系的步骤包括原生质体的制备、转化与再生, 其中关键是原生质体的制备, 通常采用酶解法破除细胞壁获得原生质体, 其获得率直接影响转化效率。转化通常借助于聚乙二醇(PEG)介导、电击或限制性内切酶法完成, 其中PEG介导的原生质体转化法较传统, 技术较成熟; 电击法操作较简单; 限制性内切酶(REMI)介导的整合转化, 因其转化效率较高, 可产生大量稳定的转化子, 正被越来越多地应用于丝状真菌的遗传转化。 |
| 英文摘要: |
| The use of protoplast-mediated genetic transformation system is one of the means to achieve large-scale randomized and site-specific mutagenesis as well as strain improvement in fungi. It lays a foundation for studying fungal pathogenicity and infection mechanism. The procedure of constructing a protoplast-mediated transformation system involves protoplast preparation, transformation and regeneration. Among them, protoplast preparation is the key step, which is usually obtained by enzyme digestion-based cell wall removal. Protoplast production rate directly affects transformation efficiency. Transformation is usually mediated by polyethylene glycol (PEG), electric shock or restriction endonuclease digestion; PEG-mediated protoplast transformation is a relatively traditional and mature technique. The electric shock method is featured by simple operation. Because of the higher transformation efficiency which can produce a large number of stable transformants, restriction enzyme mediated integration (REMI) method is increasingly used in the genetic transformation of filamentous fungi. |
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