DC Field | Value | Language |
---|---|---|
dc.contributor.author | Seo, Hogyu David | ko |
dc.contributor.author | Lee, Daeyoup | ko |
dc.date.accessioned | 2018-10-19T00:33:05Z | - |
dc.date.available | 2018-10-19T00:33:05Z | - |
dc.date.created | 2018-09-27 | - |
dc.date.created | 2018-09-27 | - |
dc.date.issued | 2018-05 | - |
dc.identifier.citation | JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, no.135 | - |
dc.identifier.issn | 1940-087X | - |
dc.identifier.uri | http://hdl.handle.net/10203/245944 | - |
dc.description.abstract | Random mutagenesis of a target gene is commonly used to identify mutations that yield the desired phenotype. Of the methods that may be used to achieve random mutagenesis, error-prone PCR is a convenient and efficient strategy for generating a diverse pool of mutants (i.e., a mutant library). Error-prone PCR is the method of choice when a researcher seeks to mutate a pre-defined region, such as the coding region of a gene while leaving other genomic regions unaffected. After the mutant library is amplified by error-prone PCR, it must be cloned into a suitable plasmid. The size of the library generated by error-prone PCR is constrained by the efficiency of the cloning step. However, in the fission yeast, Schizosaccharomyces pombe, the cloning step can be replaced by the use of a highly efficient one-step fusion PCR to generate constructs for transformation. Mutants of desired phenotypes may then be selected using appropriate reporters. Here, we describe this strategy in detail, taking as an example, a reporter inserted at centromeric heterochromatin. | - |
dc.language | English | - |
dc.publisher | JOURNAL OF VISUALIZED EXPERIMENTS | - |
dc.subject | IN-VITRO | - |
dc.subject | TRANSCRIPTION | - |
dc.subject | PCR | - |
dc.subject | ACTIVATION | - |
dc.title | Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast | - |
dc.type | Article | - |
dc.identifier.wosid | 000444372200086 | - |
dc.identifier.scopusid | 2-s2.0-85050659633 | - |
dc.type.rims | ART | - |
dc.citation.issue | 135 | - |
dc.citation.publicationname | JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | - |
dc.identifier.doi | 10.3791/57499 | - |
dc.contributor.localauthor | Lee, Daeyoup | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Genetics | - |
dc.subject.keywordAuthor | Issue 135 | - |
dc.subject.keywordAuthor | Random mutagenesis | - |
dc.subject.keywordAuthor | fission yeast | - |
dc.subject.keywordAuthor | heterochromatin | - |
dc.subject.keywordAuthor | error-prone PCR | - |
dc.subject.keywordAuthor | centromere | - |
dc.subject.keywordAuthor | gene targeting | - |
dc.subject.keywordPlus | IN-VITRO | - |
dc.subject.keywordPlus | TRANSCRIPTION | - |
dc.subject.keywordPlus | PCR | - |
dc.subject.keywordPlus | ACTIVATION | - |
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