Genetic engineering system for syngas-utilizing acetogen, Eubacterium limosum KIST612

Bioresource Technology Reports ◽

10.1016/j.biteb.2020.100452 ◽

2020 ◽

Vol 11 ◽

pp. 100452 ◽

Cited By ~ 1

Author(s):

Jiyeong Jeong ◽

Ji-Yeon Kim ◽

Byeonghyeok Park ◽

In-Geol Choi ◽

In Seop Chang

Keyword(s):

Genetic Engineering ◽

Engineering System ◽

Eubacterium Limosum

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Genetic engineering of marine cyanophages reveals integration but not lysogeny in T7-like cyanophages

The ISME Journal ◽

10.1038/s41396-021-01085-8 ◽

2021 ◽

Author(s):

Dror Shitrit ◽

Thomas Hackl ◽

Raphael Laurenceau ◽

Nicolas Raho ◽

Michael C. G. Carlson ◽

...

Keyword(s):

Genetic Engineering ◽

Attachment Site ◽

Marine Cyanobacteria ◽

Engineering System ◽

Pcr Screening ◽

Photosynthetic Organisms ◽

Infection Dynamics ◽

Attachment Sites ◽

Insight Into

AbstractMarine cyanobacteria of the genera Synechococcus and Prochlorococcus are the most abundant photosynthetic organisms on earth, spanning vast regions of the oceans and contributing significantly to global primary production. Their viruses (cyanophages) greatly influence cyanobacterial ecology and evolution. Although many cyanophage genomes have been sequenced, insight into the functional role of cyanophage genes is limited by the lack of a cyanophage genetic engineering system. Here, we describe a simple, generalizable method for genetic engineering of cyanophages from multiple families, that we named REEP for REcombination, Enrichment and PCR screening. This method enables direct investigation of key cyanophage genes, and its simplicity makes it adaptable to other ecologically relevant host-virus systems. T7-like cyanophages often carry integrase genes and attachment sites, yet exhibit lytic infection dynamics. Here, using REEP, we investigated their ability to integrate and maintain a lysogenic life cycle. We found that these cyanophages integrate into the host genome and that the integrase and attachment site are required for integration. However, stable lysogens did not form. The frequency of integration was found to be low in both lab cultures and the oceans. These findings suggest that T7-like cyanophage integration is transient and is not part of a classical lysogenic cycle.

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