The Synthesis and Assembly of a Truncated Cyanophage Genome and Its Expression in a Heterogenous Host.
The Synthesis and Assembly of a Truncated Cyanophage Genome and Its Expression in a Heterogenous Host.
复制标题
截短的噬藻体基因组的合成和组装及其在异源宿主中的表达
DOI：
10.3390/life12081234
复制
发表时间：
2022-08-15
期刊：
影响因子：
3.2
通讯作者：
Jiang, Jianlan
中科院分区：
文献类型：
--
作者：
Liu, Shujing;Feng, Jia;Sun, Tao;Xu, Bonan;Zhang, Jiabao;Li, Guorui;Zhou, Jianting;Jiang, Jianlan
关键词：
噬藻体在调节水圈中蓝藻群落的动态方面发挥着重要作用，是一种很有前途的蓝藻水华生物控制策略。然而，大多数噬藻体是宿主特异性的，使得难以通过单个或多个噬藻体控制水华蓝藻。为了解决这个问题，我们探索噬藻体和它们的异源宿主之间的相互作用，目的是揭示设计和构建针对多种蓝藻宿主的人工噬藻体基因组的原则。在本研究中，我们使用合成的生物学方法来评估在各种环境条件下将噬藻体基因组片段引入异源蓝藻的影响。以天然噬藻体A-4L基因组（41，750 bp）为基础，在酿酒酵母中合成并组装了截短的噬藻体基因组Syn-A-4-8。我们发现A-4L基因组的351- 15，930 bp区域具有在尝试组装全长A-4L基因组的过程中对大肠杆菌致命的片段。将Syn-A-4-8成功地导入E. coli中，然后通过接合转移到模式蓝细菌细长聚球藻PCC 7942（Syn 7942）中。虽然在正常条件下未观察到携带Syn-A-4-8（LS-02）的Syn 7942的显著表型，但在290毫摩尔NaCl胁迫下，与对照菌株相比，其生长表现出延长的滞后期。最后，通过比较转录组学研究揭示了LS-02耐盐性改变的机制，Syn-A-4-8上的ORF 25和ORF 26是导致耐盐性改变的关键基因。我们的研究代表了设计人工噬藻体对多个主机的重要尝试，并提供了新的未来的见解蓝藻水华的控制。
Cyanophages play an important role in regulating the dynamics of cyanobacteria communities in the hydrosphere, representing a promising biological control strategy for cyanobacterial blooms. Nevertheless, most cyanophages are host-specific, making it difficult to control blooming cyanobacteria via single or multiple cyanophages. In order to address the issue, we explore the interaction between cyanophages and their heterologous hosts, with the aim of revealing the principles of designing and constructing an artificial cyanophage genome towards multiple cyanobacterial hosts. In the present study, we use synthetic biological approaches to assess the impact of introducing a fragment of cyanophage genome into a heterologous cyanobacterium under a variety of environmental conditions. Based on a natural cyanophage A-4L genome (41,750 bp), a truncated cyanophage genome Syn-A-4-8 is synthesized and assembled in Saccharomyces cerevisiae. We found that a 351–15,930 bp area of the A-4L genome has a fragment that is lethal to Escherichia coli during the process of attempting to assemble the full-length A-4L genome. Syn-A-4-8 was successfully introduced into E. coli and then transferred into the model cyanobacterium Synechococcus elongatus PCC 7942 (Syn7942) via conjugation. Although no significant phenotypes of Syn7942 carrying Syn-A-4-8 (LS-02) could be observed under normal conditions, its growth exhibited a prolonged lag phase compared to that of the control strain under 290-millimolar NaCl stress. Finally, the mechanisms of altered salt tolerance in LS-02 were revealed through comparative transcriptomics, and ORF25 and ORF26 on Syn-A-4-8 turned out to be the key genes causing the phenotype. Our research represents an important attempt in designing artificial cyanophages towards multiple hosts, and offers new future insights into the control of cyanobacterial blooms.
登录
查看更多内容
影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
20.6
作者：
Chen W;Han M;Zhou J;Ge Q;Wang P;Zhang X;Zhu S;Song L;Yuan Y
通讯作者：
Yuan Y
影响因子：
5.1
作者：
Dekel-Bird, Naama P.;Sabehi, Gazalah;Lindell, Debbie
通讯作者：
Lindell, Debbie
影响因子：
5.2
作者：
Hanson CA;Marston MF;Martiny JB
通讯作者：
Martiny JB