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Published online on January 3, 2006, 10.1073/pnas.0510013103
PNAS | January 10, 2006 | vol. 103 | no. 2 | 425-430
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MICROBIOLOGY
Essential genes of a minimal bacterium

John I. Glass, Nacyra Assad-Garcia, Nina Alperovich, Shibu Yooseph, Matthew R. Lewis, Mahir Maruf, Clyde A. Hutchison, III, Hamilton O. Smith *, and J. Craig Venter

Synthetic Biology Group, J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850

Contributed by Hamilton O. Smith, November 18, 2005

Mycoplasma genitalium has the smallest genome of any organism that can be grown in pure culture. It has a minimal metabolism and little genomic redundancy. Consequently, its genome is expected to be a close approximation to the minimal set of genes needed to sustain bacterial life. Using global transposon mutagenesis, we isolated and characterized gene disruption mutants for 100 different nonessential protein-coding genes. None of the 43 RNA-coding genes were disrupted. Herein, we identify 382 of the 482 M. genitalium protein-coding genes as essential, plus five sets of disrupted genes that encode proteins with potentially redundant essential functions, such as phosphate transport. Genes encoding proteins of unknown function constitute 28% of the essential protein-coding genes set. Disruption of some genes accelerated M. genitalium growth.

minimal cell | Mycoplasma genitalium | synthetic biology | transposon mutagenesis

Author contributions: J.I.G., N.A.-G., C.A.H., H.O.S., and J.C.V. designed research; J.I.G., N.A.-G., N.A., M.R.L., and M.M. performed research; J.I.G. contributed new reagents/analytic tools; J.I.G. and S.Y. analyzed data; and J.I.G., C.A.H., and H.O.S. wrote the paper.

Conflict of interest statement: No conflicts declared.

Freely available online through the PNAS open access option.

Data deposition: The sequence reported in this paper has been deposited in the GenBank database (Mycoplasma genitalium G37 ATCC 33530 genomic sequence; accession no. L43967 [GenBank] ).

* To whom correspondence should be addressed. E-mail: hsmith{at}venterinstitute.org.

© 2006 by The National Academy of Sciences of the USA


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