Anuvarsh Week 11

Burkholderia cenocepacia Genome Paper

Holden, M. T., Seth-Smith, H. M., Crossman, L. C., Sebaihia, M., Bentley, S. D., Cerdeño-Tárraga, A. M., ... & Parkhill, J. (2009). The genome of Burkholderia cenocepacia J2315, an epidemic pathogen of cystic fibrosis patients. Journal of bacteriology, 191(1), 261-277.

Outline of Paper

• Introduction
  • Burkholderia cenocepacia is a known cystic fibrosis pathogen.
    • It appeared in 1980 when CF patients experienced several symptoms that resulted in early death.
    • The resulting decline in health was then on referred to as “cepacia syndrome”.
    • The specific strain associated with the “cepacia syndrome” is the ET12 strain.
  • B. cenocepecia J2315 is part the ET12 ancestral linage.
  • The researchers sequenced the J2315 strain in order to find information regarding the how the ET12 linage adapted to result in the previously mentioned clinical symptoms.
    • The sequence of the J2315 strain would also give insight on the performance of the J2315 strain in patients with CF.
• Materials and Methods
  • Bacterial strains
    • B. cenocepacia was first isolated in 1989 in the UK.
    • J2315 is resistant to several different drugs
    • In addition to J2315, 7 other previously sequenced strains were also used in this study.
  • Genome sequencing
    • Whole genome sequencing
      • DNA was extracted from J2315 and sequence data was obtained.
      • Sequence was annotated using the Artemis software.
      • Initial coding sequences were identified using Orpheus.
      • The CDSs were verified using protein databases in conjunction with BLAST and the DNA sequence was verified through all 6.
    • Comparative genomics
      • Comparisons of genome sequences were run using Artemis Comparison Tool
    • B. cenocepecia strain J2315 was compared to several other burkholderia species
  • PCR screening
    • CRP amplification was done using Platinum Pfx DNA polymerase with suppliers protocols.

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