Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated Recombination
De novo synthesis of phage genomes enables flexible genome modification and simplification. This study explores the synthetic genome assembly of <i>Pseudomonas</i> phage vB_PaeS_SCUT-S4 (S4), a 42,932 bp headful packaging phage, which encapsidates a terminally redundant, double-stranded...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-12-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/17/1/45 |
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| author | Cheng Lu Lan He Yangyijun Guo Tingting Wang Yanrui Ye Zhanglin Lin |
| author_facet | Cheng Lu Lan He Yangyijun Guo Tingting Wang Yanrui Ye Zhanglin Lin |
| author_sort | Cheng Lu |
| collection | DOAJ |
| description | De novo synthesis of phage genomes enables flexible genome modification and simplification. This study explores the synthetic genome assembly of <i>Pseudomonas</i> phage vB_PaeS_SCUT-S4 (S4), a 42,932 bp headful packaging phage, which encapsidates a terminally redundant, double-stranded DNA genome exceeding unit length. We demonstrate that using the yeast TAR approach, the S4 genome can be assembled and rebooted from a unit-length genome plus a minimal 60 bp terminal redundant sequence. Furthermore, we show that S4 can be synthesized from arbitrary starting nucleotides and modified with a red fluorescent protein as a reporter. Additionally, we successfully designed and assembled synthetic S4 phages with reduced genomes, knocking out up to 10 of the 24 hypothetical genes simultaneously, with a combined length of 2883 bp, representing 6.7% of the unit-length genome. This work highlights the potential for engineering simplified, customizable headful packaging phage genomes, providing a foundation for future studies of these phages for potential clinical applications. |
| format | Article |
| id | doaj-art-03b8b7f9630f4a7bba7c616d3bd12676 |
| institution | Kabale University |
| issn | 1999-4915 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj-art-03b8b7f9630f4a7bba7c616d3bd126762025-01-24T13:52:24ZengMDPI AGViruses1999-49152024-12-011714510.3390/v17010045Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated RecombinationCheng Lu0Lan He1Yangyijun Guo2Tingting Wang3Yanrui Ye4Zhanglin Lin5School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaSchool of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, ChinaDe novo synthesis of phage genomes enables flexible genome modification and simplification. This study explores the synthetic genome assembly of <i>Pseudomonas</i> phage vB_PaeS_SCUT-S4 (S4), a 42,932 bp headful packaging phage, which encapsidates a terminally redundant, double-stranded DNA genome exceeding unit length. We demonstrate that using the yeast TAR approach, the S4 genome can be assembled and rebooted from a unit-length genome plus a minimal 60 bp terminal redundant sequence. Furthermore, we show that S4 can be synthesized from arbitrary starting nucleotides and modified with a red fluorescent protein as a reporter. Additionally, we successfully designed and assembled synthetic S4 phages with reduced genomes, knocking out up to 10 of the 24 hypothetical genes simultaneously, with a combined length of 2883 bp, representing 6.7% of the unit-length genome. This work highlights the potential for engineering simplified, customizable headful packaging phage genomes, providing a foundation for future studies of these phages for potential clinical applications.https://www.mdpi.com/1999-4915/17/1/45headful packagingsynthetic phageyeast transformation-associated recombination<i>Pseudomonas aeruginosa</i> phage |
| spellingShingle | Cheng Lu Lan He Yangyijun Guo Tingting Wang Yanrui Ye Zhanglin Lin Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated Recombination Viruses headful packaging synthetic phage yeast transformation-associated recombination <i>Pseudomonas aeruginosa</i> phage |
| title | Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated Recombination |
| title_full | Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated Recombination |
| title_fullStr | Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated Recombination |
| title_full_unstemmed | Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated Recombination |
| title_short | Synthesis of Headful Packaging Phages Through Yeast Transformation-Associated Recombination |
| title_sort | synthesis of headful packaging phages through yeast transformation associated recombination |
| topic | headful packaging synthetic phage yeast transformation-associated recombination <i>Pseudomonas aeruginosa</i> phage |
| url | https://www.mdpi.com/1999-4915/17/1/45 |
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