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Comprehensive Identification of Conditionally Essential Genes in Mycobacteria

Comprehensive Identification of Conditionally Essential Genes in Mycobacteria. Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12712-7. Epub 2001 Oct 16. How to Determine the Functions of Genes?. DNA microarray PCR based mutagenesis Signature tagged mutagenesis (STM)

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Comprehensive Identification of Conditionally Essential Genes in Mycobacteria

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  1. Comprehensive Identification of Conditionally Essential Genes in Mycobacteria Proc Natl Acad Sci U S A. 2001 Oct 23;98(22):12712-7. Epub 2001 Oct 16.

  2. How to Determine the Functions of Genes? DNA microarray PCR based mutagenesis Signature tagged mutagenesis (STM) Genomic Analysis and Mapping By in vitro Transposition (GAMBIT)

  3. Nature Reviews Genetics 7:929-939 (2006)

  4. Signature Tagged Mutagenesis (STM) Nature Reviews Genetics 7:929-939 (2006)

  5. Genomic Analysis and Mapping By in vitro Transposition (GAMBIT) Annual Review of Microbiology53:129-54 (1999)

  6. Nature Reviews Genetics 7:929-939 (2006)

  7. Generation of Mutant Libraries

  8. Labeled RNA Synthesis

  9. Transposon-Site Hybridization Rich medium Minimal medium

  10. Minimal medium Rich medium

  11. Auxotrophic Mutations Identified by TraSH Rich medium Minimal medium Ratio (Rich/Minimal) Gene

  12. Superpathway of Leu, Val, and Ile biosynthesis ilvA leuD leuB BioCyc

  13. Superpathway of sulfate assimilation and Cysbiosynthesis cysD cysH BioCyc

  14. EMBO Journal15:5470-5479 (1996)

  15. Codon Usage of M. bovis • TAT/TAC Tyr 2.06% (100% of Tyr) • TTA/CTA Leu 0.63% (6.4% of Leu) • ATA Ile 0.22% (5.2% of Ile) • GTA Val 0.48% (5.6% of Val) • TAA/TAG Stop 0.14% (46.6% of Stop) • TATAAT -10 element CBS

  16. Thanks for your attention!

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