Construction of a Bacterial Cell that Contains Only the Set of Essential Genes Necessary to Impart Life
R&D Statys reot, 17 Aug-16 Nov 2013
J CRAIG VENTER INST ROCKVILLE MD
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We have made significant progress on all fronts of the project. On the Top Down approach to genome minimization, we have used the Essential E, Non-essential N, and Impaired I gene categories to make steady progress with gene and gene cluster deletions. To date, we have removed approximately 273 kb from the Mycoplasma mycoides JCVI-syn1.0 genome. The resultant 806 kb genome is viable and grows with a normal growth rate. Results on the Bottom Up approach are substantial. We have continued work on the reduced genome design RGD1 based on the NEI categorization system and completed testing on 6 out of 8 segments from the RGD1 design. All six segments are viable in a 78th wild type background. In separate experiments, we have also tested genomes containing 2 Segments 26 or 3 Segments 268 minimized segments. These two genome are also viable. The effort to modularize the genome is in progress. A tRNA module was constructed and inserted into the M. mycoides syn1.0 genome. The resulting genome is viable following transplantation. Preliminary work aimed at genome complementation has continued. Experiments using a transposon based approach have been successful, and allowed recovery of growth rate on a genome with an impaired growth rate.
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