Building artificial genetic circuits to understand protein function
Intrinsic protein properties that may not be apparent by only examining three-dimensional structures can be revealed by careful analysis of mutant protein variants. Deep mutational scanning is a technique that allows the functional analysis of millions of protein variants in a single experiment. To...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Book Chapter |
| Language: | English |
| Published: |
ACADEMIC PRESS LTD-ELSEVIER SCIENCE LTD
2020
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| Subjects: | |
| Online Access: | https://research-repository.uwa.edu.au/files/81276020/Scott_et_al._Author_Manuscript.pdf http://hdl.handle.net/20.500.11937/90960 |
| _version_ | 1848765472603897856 |
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| author | Scott, L.H. Mathews, J.C. Filipovska, A. Rackham, Oliver |
| author2 | Shukla, AK |
| author_facet | Shukla, AK Scott, L.H. Mathews, J.C. Filipovska, A. Rackham, Oliver |
| author_sort | Scott, L.H. |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | Intrinsic protein properties that may not be apparent by only examining three-dimensional structures can be revealed by careful analysis of mutant protein variants. Deep mutational scanning is a technique that allows the functional analysis of millions of protein variants in a single experiment. To enable this high-throughput technique, the mutant genotype of protein variants must be coupled to a selectable function. This chapter outlines how artificial genetic circuits in the yeast Saccharomyces cerevisiae can maintain the genotype-phenotype link, thus enabling the general application of this approach. To do this, we describe how to engineer genetic selections in yeast, methods to construct mutant libraries, and how to analyze sequencing data. We investigate the structure-function relationships of the antimicrobial resistance protein TetX to illustrate this process. In doing so, we demonstrate that deep mutational scanning is a powerful method to dissect the importance of individual residues for the inactivation of antibiotic analogues, with consequences for the rational design of new drugs to combat antimicrobial resistance. |
| first_indexed | 2025-11-14T11:35:48Z |
| format | Book Chapter |
| id | curtin-20.500.11937-90960 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T11:35:48Z |
| publishDate | 2020 |
| publisher | ACADEMIC PRESS LTD-ELSEVIER SCIENCE LTD |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-909602023-06-13T08:47:36Z Building artificial genetic circuits to understand protein function Scott, L.H. Mathews, J.C. Filipovska, A. Rackham, Oliver Shukla, AK Science & Technology Life Sciences & Biomedicine Biochemical Research Methods Biochemistry & Molecular Biology Cell Biology YEAST RESISTANCE PARALLEL TOOLKIT SYSTEM TRANSFORMATION RECOGNITION TIGECYCLINE EXPRESSION EFFICIENCY Antibiotic resistance Biosensor Deep mutational scanning Genetic circuit Structure-function relationship Synthetic biology Gene Regulatory Networks Mutant Proteins Mutation Proteins Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Mutation Mutant Proteins Gene Regulatory Networks Intrinsic protein properties that may not be apparent by only examining three-dimensional structures can be revealed by careful analysis of mutant protein variants. Deep mutational scanning is a technique that allows the functional analysis of millions of protein variants in a single experiment. To enable this high-throughput technique, the mutant genotype of protein variants must be coupled to a selectable function. This chapter outlines how artificial genetic circuits in the yeast Saccharomyces cerevisiae can maintain the genotype-phenotype link, thus enabling the general application of this approach. To do this, we describe how to engineer genetic selections in yeast, methods to construct mutant libraries, and how to analyze sequencing data. We investigate the structure-function relationships of the antimicrobial resistance protein TetX to illustrate this process. In doing so, we demonstrate that deep mutational scanning is a powerful method to dissect the importance of individual residues for the inactivation of antibiotic analogues, with consequences for the rational design of new drugs to combat antimicrobial resistance. 2020 Book Chapter http://hdl.handle.net/20.500.11937/90960 10.1016/bs.mie.2019.11.003 English https://research-repository.uwa.edu.au/files/81276020/Scott_et_al._Author_Manuscript.pdf http://purl.org/au-research/grants/arc/DP180101656 ACADEMIC PRESS LTD-ELSEVIER SCIENCE LTD restricted |
| spellingShingle | Science & Technology Life Sciences & Biomedicine Biochemical Research Methods Biochemistry & Molecular Biology Cell Biology YEAST RESISTANCE PARALLEL TOOLKIT SYSTEM TRANSFORMATION RECOGNITION TIGECYCLINE EXPRESSION EFFICIENCY Antibiotic resistance Biosensor Deep mutational scanning Genetic circuit Structure-function relationship Synthetic biology Gene Regulatory Networks Mutant Proteins Mutation Proteins Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Mutation Mutant Proteins Gene Regulatory Networks Scott, L.H. Mathews, J.C. Filipovska, A. Rackham, Oliver Building artificial genetic circuits to understand protein function |
| title | Building artificial genetic circuits to understand protein function |
| title_full | Building artificial genetic circuits to understand protein function |
| title_fullStr | Building artificial genetic circuits to understand protein function |
| title_full_unstemmed | Building artificial genetic circuits to understand protein function |
| title_short | Building artificial genetic circuits to understand protein function |
| title_sort | building artificial genetic circuits to understand protein function |
| topic | Science & Technology Life Sciences & Biomedicine Biochemical Research Methods Biochemistry & Molecular Biology Cell Biology YEAST RESISTANCE PARALLEL TOOLKIT SYSTEM TRANSFORMATION RECOGNITION TIGECYCLINE EXPRESSION EFFICIENCY Antibiotic resistance Biosensor Deep mutational scanning Genetic circuit Structure-function relationship Synthetic biology Gene Regulatory Networks Mutant Proteins Mutation Proteins Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Mutation Mutant Proteins Gene Regulatory Networks |
| url | https://research-repository.uwa.edu.au/files/81276020/Scott_et_al._Author_Manuscript.pdf https://research-repository.uwa.edu.au/files/81276020/Scott_et_al._Author_Manuscript.pdf http://hdl.handle.net/20.500.11937/90960 |