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1.6K Views
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13:22 min
September 13th, 2022
DOI :
10.3791/63967-v
Chapters
0:04
Introduction
1:00
Transforming cDNA Library into the QuerySchizosaccharomyces pombe Mutant Strain and Screening Multicopy Suppressors
5:16
Validating the Suppressors of the Phenotype Associated with the Query Mutant Strain
7:41
Isolation of the Plasmid from the Schizosaccharomyces pombeSuppressor Clones
9:55
Identification of the Gene Encoded by the Suppressor Clone
10:48
Results: Screening Multicopy Suppressor of ell1 Deletion-Associated Genotoxic Stress Sensitivity
12:49
Conclusion
Transcript
抑制因子筛选识别遗传相互作用,这些相互作用将揭示目的基因的功能以及它们可能参与体内的途径和生物过程。该技术可以识别两种基因产物之间的遗传关系,而这两种基因产物可能尚未使用其他方法证明。酵母中这种筛选的结果可以扩展到高真核生物,因为大多数基本的生物途径和过程在整个进化过程中都是保守的。
该方案的成功取决于其转化为酵母细胞的高质量和高效率的可用性。因此,在YE培养基板上以32摄氏度培养裂殖酵母菌绒毛1缺失菌株之前,质粒的转化方案。从平板中取出一个充满ell1突变菌株的环,接种5毫升
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Summary
这项工作描述了 在Pombe裂殖酵母中进行多拷贝抑制遗传筛选的方案。该筛选使用全基因组质粒文库来鉴定与查询突变菌株相关的功能丧失表型的抑制克隆。使用该筛选鉴定 ell1 零突变体的新型遗传抑制因子。
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