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Polar Effects of Transposon Insertion into a Minimal Bacterial Genome
Abstract
In studies of the minimal genetic requirements for life, we used global transposon mutagenesis to identify genes needed for a minimal bacterial genome. Transposon insertion can disrupt the function of a gene but can also have polar effects on the expression of adjacent genes. In the Tn 5 -Puro r construct used in our studies, read-through transcription from Tn 5 -Puro r can drive expression of downstream genes. This results in a preference for Tn 5 -Puro r insertions transcribed toward a downstream essential or quasi-essential gene within the same operon. Such polar effects can have an impact on the classification of genes as essential, quasi-essential, or nonessential, but this has been observed in only a few cases. Also, polar effects of Tn 5 -Puro r insertion can sometimes give clues to gene function.
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“we constructed in‐frame, scar‐less deletion mutants of P. putida KT2440—to circumvent any potential polar effect brought about by transposon insertion (Hutchison et al., 2019).”
From Role of the CrcB transporter of Pseudomonas putida in the multi‐level stress response elicited by mineral fluoride (2022) · cites this paper as Hutchison et al., 2019
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Design and synthesis of a minimal bacterial genome
Hutchison, C. A., Chuang, R.-Y., Noskov, V. N., Assad-Garcia, N., Deerinck, T. J., Ellisman, M. H., Gill, J., Kannan, K., Karas, B. J., Ma, L., et al. (2016) · Science · doi:10.1126/science.aad6253 · Cited by 1,500 · cited 4×
“Junctions between Tn5-Puror and the genome were amplified by an inverse PCR strategy and were subjected to deep sequencing (2).”
BackgroundMethodInfluentialSelf-citationEssential metabolism for a minimal cell
Breuer, M., Earnest, E. E., Merryman, C., Wise, K. S., Sun, L., Lynott, M. R., Hutchison, C. A., Smith, H. O., Lapek, J. D., Gonzalez, D. J., et al. (2019) · eLife · doi:10.7554/elife.36842 · Cited by 190 · cited 2×
“Gene _0132, genes _0132 and _0133, and gene _0728 were each knocked out from the syn3A genome (9) carried in yeast strain VL6-48N-Cas9 (a gift from Daniel G.”
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Research works printed but not linked: 11
- 10.1126/science.286.5447.2165 · DOI
- 10.1016/0092-8674(80)90305-0 · DOI
- 10.1038/nmeth.1377 · DOI
- 10.1128/AAC.00388-09 · DOI
- 10.1007/s00284-009-9455-2 · DOI
- 10.1016/0022-2836(92)90574-4 · DOI
- 10.1128/JB.00124-09 · DOI
- 10.1016/j.molcel.2018.01.003 · DOI
- 10.1371/journal.pcbi.1006946 · DOI
- 10.1126/science.1190719 · DOI · Paper page
- 10.1038/srep30714 · DOI
Cited by
Sorted by citations
Calero, P., Gurdo, N., & Nikel, P. I. (2022) · Environmental Microbiology · doi:10.1111/1462-2920.16110 · Cited by 56
Metadata
- Authors
- Hutchison, C. A., Merryman, C., Sun, L., Assad-Garcia, N., Richter, R. A., Smith, H. O., & Glass, J. I.
- Affiliations
- Synthetic Biology Group, J. Craig Venter Institute, La Jolla, California, USA
- Synthetic Biology Group, J. Craig Venter Institute, Rockville, Maryland, USA
- Venue
- Journal of Bacteriology
- Publisher
- American Society for Microbiology
- Published
- 2019-10
- Volume
- 201
- Issue
- 19
- ISSN
- 0021-9193
- 1098-5530
- PMC
- PMC6755753
- Cited by
- 47
- References
- 2
- Language
- en
- Funders
- US National Science Foundation (MCB-1840320, MCB-1818344)
- J. Craig Venter Institute
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