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Science 10 December 1999:
Vol. 286. no. 5447, pp. 2165 - 2169
DOI: 10.1126/science.286.5447.2165
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Reports

Global Transposon Mutagenesis and a Minimal Mycoplasma Genome

Clyde A. Hutchison III, 12* Scott N. Peterson, 1*dagger Steven R. Gill, 1 Robin T. Cline, 1 Owen White, 1 Claire M. Fraser, 1 Hamilton O. Smith, 1ddagger J. Craig Venter 1ddagger §

Mycoplasma genitalium with 517 genes has the smallest gene complement of any independently replicating cell so far identified. Global transposon mutagenesis was used to identify nonessential genes in an effort to learn whether the naturally occurring gene complement is a true minimal genome under laboratory growth conditions. The positions of 2209 transposon insertions in the completely sequenced genomes of M. genitalium and its close relative M. pneumoniae were determined by sequencing across the junction of the transposon and the genomic DNA. These junctions defined 1354 distinct sites of insertion that were not lethal. The analysis suggests that 265 to 350 of the 480 protein-coding genes of M. genitalium are essential under laboratory growth conditions, including about 100 genes of unknown function.

1 The Institute for Genomic Research, 9712 Medical Center Drive, Rockville, MD 20850, USA.
2 Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
*   These authors contributed equally to this report.

dagger    To whom reprint requests should be addressed.

ddagger    Present address: Celera Genomics, 45 West Gude Drive, Rockville, MD 20850, USA.

§   To whom correspondence should be addressed.

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