Design and synthesis of a minimal bacterial genome

Published

March 25, 2016

Author(s)

Elizabeth A. Strychalski, Clyde A. Hutchinson III, Ray-Yuan Chuang, Vladimir Noskov, Nacyra Assad-Garcia, Tom J. Deerinck, Mark H. Ellisman, John Gill, Krishna Kannan, Bogumil J. Karas, Li Ma, James F. Pelletier, Zhi-Qing Qi, Alexander Richter, Lijie Sun, Yo Suzuki, Billyana Tsvetanova, Kim S. Wise, Hamilton O. Smith, John I. Glass, Chuck Merryman, Daniel G. Gibson, J. C. Venter

Abstract

We used whole-genome design and complete chemical synthesis to minimize the 1079–kilobase pair synthetic genome of Mycoplasma mycoides JCVI-syn1.0. An initial design, based on collective knowledge of molecular biology combined with limited transposon mutagenesis data, failed to produce a viable cell. Improved transposon mutagenesis methods revealed a class of quasi- essential genes that are needed for robust growth, explaining the failure of our initial design. Three cycles of design, synthesis, and testing, with retention of quasi-essential genes, produced JCVI-syn3.0 (531 kilobase pairs, 473 genes), which has a genome smaller than that of any autonomously replicating cell found in nature. JCVI-syn3.0 retains almost all genes involved in the synthesis and processing of macromolecules. Unexpectedly, it also contains 149 genes with unknown biological functions. JCVI-syn3.0 is a versatile platform for investigating the core functions of life and for exploring whole-genome design.

Citation

Science

Volume

351

Issue

6280

Pub Type

Journals

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DOI Link

Citation

Strychalski, E. , Hutchinson, C. , Chuang, R. , Noskov, V. , Assad-Garcia, N. , Deerinck, T. , Ellisman, M. , Gill, J. , Kannan, K. , Karas, B. , Ma, L. , Pelletier, J. , Qi, Z. , Richter, A. , Sun, L. , Suzuki, Y. , Tsvetanova, B. , Wise, K. , Smith, H. , Glass, J. , Merryman, C. , Gibson, D. and Venter, J. (2016), Design and synthesis of a minimal bacterial genome, Science, [online], https://doi.org/10.1126/ science.aad6253 (Accessed April 30, 2024)