كتاب
Genetic Transformation of Candida auris via Homology-Directed Repair Using a Standard Lithium Acetate Protocol
| العنوان: | Genetic Transformation of Candida auris via Homology-Directed Repair Using a Standard Lithium Acetate Protocol |
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| المؤلفون: | Bravo Ruiz, Gustavo, Lorenz, Alexander |
| المساهمون: | Lorenz, Alexander |
| المصدر: | Bravo Ruiz , G & Lorenz , A 2022 , Genetic Transformation of Candida auris via Homology-Directed Repair Using a Standard Lithium Acetate Protocol . in A Lorenz (ed.) , Candida auris : Methods and Protocols . 1 edn , Methods in Molecular Biology , vol. 2517 , Springer , pp. 95-110 . https://doi.org/10.1007/978-1-0716-2417-3_8Test |
| بيانات النشر: | Springer |
| سنة النشر: | 2022 |
| المجموعة: | Discovery - University of Dundee Online Publications |
| مصطلحات موضوعية: | Acetates, Antifungal Agents/metabolism, Candida/genetics, Candida auris, Polymerase Chain Reaction/methods, Transformation, Genetic, Gene targeting, Genetic transformation, Genetic manipulation, Dominant drug resistance markers, Gene deletion, /dk/atira/pure/subjectarea/asjc/1300/1311, name=Genetics, /dk/atira/pure/subjectarea/asjc/1300/1312, name=Molecular Biology |
| الوصف: | Reverse genetics is a particularly powerful tool in non-model organisms with known whole-genome sequences enabling the characterization of gene and, thus, protein function via a mutant phenotype. Reverse genetic approaches require genetic manipulation techniques which often need to be specifically developed for non-model organisms; this can be fraught with difficulties. Here, we describe a genetic transformation protocol for the recently emerged human pathogen Candida auris to target the integration of DNA constructs into genomic locations via homology-directed repair using long flanking homologous sequences (>1 kb). We detail the generation of DNA constructs for gene deletion with dominant drug resistance markers via fusion PCR, the transformation of these constructs into chemically competent C. auris cells, and the confirmation of correct integration by PCR. This strategy can be adapted to deliver DNA constructs other than deletion cassettes, including promoter exchanges and protein tags. |
| نوع الوثيقة: | book part |
| اللغة: | English |
| ردمك: | 978-1-07-162416-6 1-07-162416-4 |
| العلاقة: | https://discovery.dundee.ac.uk/en/publications/ec697ccf-fd59-4ca7-a3fa-4732ccf03cadTest; urn:ISBN:9781071624166 |
| DOI: | 10.1007/978-1-0716-2417-3_8 |
| الإتاحة: | https://doi.org/10.1007/978-1-0716-2417-3_8Test https://discovery.dundee.ac.uk/en/publications/ec697ccf-fd59-4ca7-a3fa-4732ccf03cadTest |
| حقوق: | info:eu-repo/semantics/closedAccess |
| رقم الانضمام: | edsbas.335F2E67 |
| قاعدة البيانات: | BASE |
| ردمك: | 9781071624166 1071624164 |
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| DOI: | 10.1007/978-1-0716-2417-3_8 |