Outpacing E. coli : Development of Vibrio natriegens as a Next-Generation Cloning Host
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Abstract
Despite transformative advances in DNA synthesis, sequencing, and automation that have accelerated recombinant DNA workflows, molecular cloning hosts have scarcely evolved past the Escherichia coli strains adopted out of convenience in the 1970s. We present NBx CyClone™ – an engineered strain of Vibrio natriegens – as a next-generation host for molecular cloning. This non-pathogenic marine bacterium combines broad plasmid and genetic tool compatibility, a versatile metabolism, and the fastest known doubling time of any free-living organism. By shortening growth-dependent steps, this host offers a practical route to faster, more efficient recombinant DNA workflows across research and industry.
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Outpacing E. coli: Development of Vibrio natriegens as a Next-Generation Cloning Host
I’m excited to see the development of a commercial V. natriegens for the community to try out. I had a few questions arise when reading your paper.
First, in the introduction section you mention “This can be scaled from small culture tubes in the laboratory (to generate microgram quantities of DNA), to large-scale industrial bioreactors (to generate kilograms of DNA for vaccine production).” I am wondering where you envision the biggest benefit occurring? Would it be mostly for commercial DNA manufacturing? For general cloning in a research lab, I’m not sure how much I’ve ever worried about E.coli’s slower doubling time for my typical cloning workflow, since I often start cultures in the evening and let them grow overnight. With this system, I’m …
Outpacing E. coli: Development of Vibrio natriegens as a Next-Generation Cloning Host
I’m excited to see the development of a commercial V. natriegens for the community to try out. I had a few questions arise when reading your paper.
First, in the introduction section you mention “This can be scaled from small culture tubes in the laboratory (to generate microgram quantities of DNA), to large-scale industrial bioreactors (to generate kilograms of DNA for vaccine production).” I am wondering where you envision the biggest benefit occurring? Would it be mostly for commercial DNA manufacturing? For general cloning in a research lab, I’m not sure how much I’ve ever worried about E.coli’s slower doubling time for my typical cloning workflow, since I often start cultures in the evening and let them grow overnight. With this system, I’m envisioning you would more likely to be starting cultures in the morning and harvesting in the afternoon. It would be nice to see a figure demonstrating what some hypothetical cloning workflows (with timelines) would look like to have a better understanding of how much time could be saved.
Second, I am wondering if you tested transformation of any larger plasmids. Most plasmids you tested ranged from ~3kb to 6kb. However, some of the larger plasmids seem like they may have a growth penalty (Figure 6) and a decrease in transformation efficiency (Fig 9). Would these possible phenotypes be more pronounced if you are transforming a plasmid that is 20-25kb, which is about the max that E.coli can support?
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