التفاصيل البيبلوغرافية
| العنوان: |
Reduction of the Bacterial Genome by Transposon-Mediated Random Deletion |
| المؤلفون: |
Shuai Ma (414625), Tianyuan Su (3454208), Jinming Liu (124265), Xuemei Lu (217979), Qingsheng Qi (256048) |
| سنة النشر: |
2022 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Microbiology, Genetics, Cancer, Space Science, Biological Sciences not elsewhere classified, rational design encounter, least 2 magnitudes, intracellular alternative end, electroporation efficiency increased, complex gene functions, allows genomic deletions, random genomic deletions, mediated random deletion, named “ transposon, five reduction cycles, random positions, widely applied, various sizes, using e, synthetic biology, strand breaks, small e, original strain, minimal genomes, metabolic engineering, limited knowledge, joining mechanism, important strategy, growth rates, escherichia coli |
| الوصف: |
Genome reduction is an important strategy in synthetic biology for constructing functional chassis cells or minimal genomes. However, the limited knowledge of complex gene functions and interactions makes genome reduction by rational design encounter a bottleneck. Here, we present an iterative and random genome reduction method for Escherichia coli, named “transposon-mediated random deletion (TMRD)”. TMRD generates random double-strand breaks (DSBs) in the genome by combining Tn 5 transposition with the CRISPR/Cas9 system and allows genomic deletions of various sizes at random positions during DSB repair through the intracellular alternative end-joining mechanism. Using E. coli MG1655 as the original strain, a pool of cells with multiple random genomic deletions were obtained after five reduction cycles. The growth rates of the obtained cells were comparable to that of MG1655, while the electroporation efficiency increased by at least 2 magnitudes. TMRD can generate a small E. coli library carrying multiple and random genomic deletions while enriching the cells with environmental fitness in the population. TMRD has the potential to be widely applied in the construction of minimal genomes or chassis cells for metabolic engineering. |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
unknown |
| Relation: |
https://figshare.com/articles/journal_contribution/Reduction_of_the_Bacterial_Genome_by_Transposon-Mediated_Random_Deletion/19104179 |
| DOI: |
10.1021/acssynbio.1c00353.s001 |
| الاتاحة: |
https://doi.org/10.1021/acssynbio.1c00353.s001 |
| Rights: |
CC BY-NC 4.0 |
| رقم الانضمام: |
edsbas.4AE9C733 |
| قاعدة البيانات: |
BASE |