Construction of a Bacterial Cell that Contains Only the Set of Essential Genes Necessary to Impart Life
Abstract
We now have a viable reduced genome design (RGD) cell that has a genome that is 85% minimized. This is a genome in which six and one half of the eight genome segments are minimized. The key issue that we must resolve to produce a fully minimized RGD is that in RGD segments 1 and 3 there are at least two instances of what we call synthetic lethal deletions that we have to identify and add back to the genome. The top down approach of iteratively removing non-essential genes from M. mycoides syn1.0 continues. Genes or sets of genes are deleted from previous top down constructs while the genomes are parked in yeast cells. Then the reduced genomes are isolated from yeast cells and transplanted to determine if the deletion is viable or has an altered phenotype. Currently we are working on our 17th iteration of this process. The smallest genome the top down effort has made to date has 764 kb and is missing 230 protein coding genes. This deletion mutant grows normally. The effort to modularize the genome is in progress on multiple fronts.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 16, 2014
- Accession Number
- ADA600795
Entities
People
- Clyde Hutchison
- Hamilton Smith
- John I Glass
- Tony Yee
Organizations
- J. Craig Venter Institute