Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene Cassettes
Genetic modification of lactic acid bacteria is an evolving and highly relevant field of research that allows the engineered bacteria to be equipped with the desired functions through the controlled expression of the recombinant protein. Novel genetic engineering techniques offer the advantage of be...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-12-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2021.797521/full |
| _version_ | 1818459867741945856 |
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| author | Tina Vida Plavec Tim Ključevšek Tim Ključevšek Aleš Berlec Aleš Berlec |
| author_facet | Tina Vida Plavec Tim Ključevšek Tim Ključevšek Aleš Berlec Aleš Berlec |
| author_sort | Tina Vida Plavec |
| collection | DOAJ |
| description | Genetic modification of lactic acid bacteria is an evolving and highly relevant field of research that allows the engineered bacteria to be equipped with the desired functions through the controlled expression of the recombinant protein. Novel genetic engineering techniques offer the advantage of being faster, easier and more efficient in incorporating modifications to the original bacterial strain. Here, we have developed a modified BglBrick system, originally introduced in Escherichia coli and optimized it for the lactic acid bacterium Lactococcus lactis. Six different expression cassettes, encoding model proteins, were assembled in different order as parts of a modified BglBrick system in a novel plasmid pNBBX. All cassettes included nisin promoter, protein encoding gene and transcription terminator. We demonstrated successful intracellular expression of the two fluorescent proteins and display of the four protein binders on the bacterial surface. These were expressed either alone or concomitantly, in combinations of three model proteins. Thus, a modified BglBrick system developed herein enables simple and modular construction of multigene plasmids and controlled simultaneous expression of three proteins in L. lactis. |
| first_indexed | 2024-12-14T23:21:11Z |
| format | Article |
| id | doaj.art-3e02a6c6ef14453bba4dd6742b03ea20 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-14T23:21:11Z |
| publishDate | 2021-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-3e02a6c6ef14453bba4dd6742b03ea202022-12-21T22:43:57ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852021-12-01910.3389/fbioe.2021.797521797521Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene CassettesTina Vida Plavec0Tim Ključevšek1Tim Ključevšek2Aleš Berlec3Aleš Berlec4Department of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaDepartment of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaFaculty of Pharmacy, University of Ljubljana, Ljubljana, SloveniaDepartment of Biotechnology, Jožef Stefan Institute, Ljubljana, SloveniaFaculty of Pharmacy, University of Ljubljana, Ljubljana, SloveniaGenetic modification of lactic acid bacteria is an evolving and highly relevant field of research that allows the engineered bacteria to be equipped with the desired functions through the controlled expression of the recombinant protein. Novel genetic engineering techniques offer the advantage of being faster, easier and more efficient in incorporating modifications to the original bacterial strain. Here, we have developed a modified BglBrick system, originally introduced in Escherichia coli and optimized it for the lactic acid bacterium Lactococcus lactis. Six different expression cassettes, encoding model proteins, were assembled in different order as parts of a modified BglBrick system in a novel plasmid pNBBX. All cassettes included nisin promoter, protein encoding gene and transcription terminator. We demonstrated successful intracellular expression of the two fluorescent proteins and display of the four protein binders on the bacterial surface. These were expressed either alone or concomitantly, in combinations of three model proteins. Thus, a modified BglBrick system developed herein enables simple and modular construction of multigene plasmids and controlled simultaneous expression of three proteins in L. lactis.https://www.frontiersin.org/articles/10.3389/fbioe.2021.797521/fullLactococcus lactisgenetic engineeringBglBrickmultifunctional bacteriaexpression system |
| spellingShingle | Tina Vida Plavec Tim Ključevšek Tim Ključevšek Aleš Berlec Aleš Berlec Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene Cassettes Frontiers in Bioengineering and Biotechnology Lactococcus lactis genetic engineering BglBrick multifunctional bacteria expression system |
| title | Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene Cassettes |
| title_full | Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene Cassettes |
| title_fullStr | Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene Cassettes |
| title_full_unstemmed | Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene Cassettes |
| title_short | Introduction of Modified BglBrick System in Lactococcus lactis for Straightforward Assembly of Multiple Gene Cassettes |
| title_sort | introduction of modified bglbrick system in lactococcus lactis for straightforward assembly of multiple gene cassettes |
| topic | Lactococcus lactis genetic engineering BglBrick multifunctional bacteria expression system |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2021.797521/full |
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