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- Tn5 Transposon-based Mutagenesis for Engineering Phage-resistant Strains of Escherichia coli BL21 (DE3)
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Yinfeng Wang , Guanhua Xuan , Houqi Ning , Jiuna Kong , Hong Lin , Jingxue Wang
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J. Microbiol. 2023;61(5):559-569. Published online May 22, 2023
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DOI: https://doi.org/10.1007/s12275-023-00048-2
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Abstract
- Escherichia coli is a preferred strain for recombinant protein production, however, it is often plagued by phage infection
during experimental studies and industrial fermentation. While the existing methods of obtaining phage-resistant strains
by natural mutation are not efficient enough and time-consuming. Herein, a high-throughput method by combining Tn5
transposon mutation and phage screening was used to produce Escherichia coli BL21 (DE3) phage-resistant strains. Mutant
strains PR281-7, PR338-8, PR339-3, PR340-8, and PR347-9 were obtained, and they could effectively resist phage infection.
Meanwhile, they had good growth ability, did not contain pseudolysogenic strains, and were controllable. The resultant
phage-resistant strains maintained the capabilities of producing recombinant proteins since no difference in mCherry red
fluorescent protein expression was found in phage-resistant strains. Comparative genomics showed that PR281-7, PR338-8,
PR339-3, and PR340-8 mutated in ecpE, nohD, nrdR, and livM genes, respectively. In this work, a strategy was successfully
developed to obtain phage-resistant strains with excellent protein expression characteristics by Tn5 transposon mutation.
This study provides a new reference to solve the phage contamination problem.
Review
- [Minireview]Potential roles of condensin in genome organization and beyond in fission yeast
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Kyoung-Dong Kim
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J. Microbiol. 2021;59(5):449-459. Published online April 20, 2021
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DOI: https://doi.org/10.1007/s12275-021-1039-2
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Abstract
- The genome is highly organized hierarchically by the function
of structural maintenance of chromosomes (SMC) complex
proteins such as condensin and cohesin from bacteria
to humans. Although the roles of SMC complex proteins have
been well characterized, their specialized roles in nuclear processes
remain unclear. Condensin and cohesin have distinct
binding sites and mediate long-range and short-range genomic
associations, respectively, to form cell cycle-specific
genome organization. Condensin can be recruited to highly
expressed genes as well as dispersed repeat genetic elements,
such as Pol III-transcribed genes, LTR retrotransposon, and
rDNA repeat. In particular, mitotic transcription factors Ace2
and Ams2 recruit condensin to their target genes, forming
centromeric clustering during mitosis. Condensin is potentially
involved in various chromosomal processes such as the
mobility of chromosomes, chromosome territories, DNA reannealing,
and transcription factories. The current knowledge
of condensin in fission yeast summarized in this review can
help us understand how condensin mediates genome organization
and participates in chromosomal processes in other
organisms.
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