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| 1种用于检测抑制基因功能的酵母株系的建立（英文） |
| Generation of a Saccharomyces cerevisiae strain suitable for testing repressor function |
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| DOI： |
| 中文关键词: 酵母 双交换插入失活 转化 |
| 英文关键词: Saccharomyces cerevisiae disruption transformation |
| 基金项目:supported by Chinese Scholarship Council and Startup Foundation for Doctors of Tibet Agricultural and Animal Husbandry College |
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| 中文摘要: |
| 酵母被广泛用于分子生物学中基因功能的检测。为扩大酵母株系UCC419在抑制基因活性检测方面的应用,本研究通过向UCC419株系中导入用特殊引物扩增出的包含标记基因TRP1的PCR片段,利用同源重组将UCC419中的筛选标记基因LEU2敲除,并同时插入TRP1,新建立的株系命名为UCC419m（m：modi-fied）。UCC419m为TRP1筛选、leu2突变型菌株,其它基因型均同UCC419。给UCC419m中转入携带LEU2的质粒pDEST32检测是否能恢复其表现型,同时转入不携带LEU2的质粒pDEST22作为阴性对照,将转化子在不含LEU2与URA3的培养基中培养,结果显示,携带LEU2质粒pDEST32的转化子能够在LEU2与URA3缺陷型培养基上正常生长,而不携带LEU2质粒pDEST22的转化子不能生长。本研究结果表明,成功建立了一种适用于基于Invitrogen载体的抑制基因活性检测或从文库中筛选抑制基因的酵母菌株。 |
| 英文摘要: |
| accharomyces cerevisiae has been widely used as tools in testing gene function.In order to utilize a Saccharomyces cerevisiae strain UCC419 for testing repressor activities,we engineered UCC419 by deleting its LEU2 gene and at the same time replacing the LEU2 with the TRP1 marker via homologous recombination.The new strain designated as UCC419m(m: modified) is now TRP1 + but leu2-.Transformation of pDEST32 that posses LEU2 into UCC419m rescued the phenotype on-Ura-Trp medium.The new strain will be useful fo... |
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