DMRT gene cluster analysis in the platypus:

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DMRT gene cluster analysis in the platypus New
insights into genomic organization and regulatory
regions
Nisrine El-Mogharbel Matthew Wakefield
Australian National University
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DMRT GENE
Members of the DSX- and
Mab-3-related transcription factor (DMRT) gene
family have been independently isolated in
vertebrates and invertebrates and are of special
interest because of their roles in sexual
differentiation.
Comparison of gene arrangement,
sequence, and expression in different
vertebrates, such as human, mouse, and puffer
fish, has been invaluable in annotating the genes
and assessing their functions
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Platypus
The egg-laying monotreme mammals
(platypus) are of particular interest in
annotating TMRT genes and assessing their
functions because of the unique phylogenetic
position they occupy at the base of the mammalian
radiation and their unique reproductive strategy.
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Materials and methods

• BAC library screening, subcloning, sequencing
• DNA preparation, tissue culture, and RNA
  isolation
• FISH
• Sequence analysis and cross-species comparisons
• Sequence analysis, Cross-species comparisons
  with VISTA Gene prediction and annotation
• Expression analysis and RT-PCR
• PCR

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Results
We isolated, sequenced,
characterized, mapped, and annotated BACs
containing platypus DMRT1, DMRT3, and DMRT2 and
compared sequences across vertebrates to identify
exons and CNGs.

• Isolation and characterization of the platypus
  DMRT gene cluster
• Prediction of DMRT genes and their putative
  products
• Colocalization of platypus DMRT1, DMRT3, and
  DMRT2

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• Sequence analysis
• Identification of CNGs
• VISTA analysis
• Repeat content analysis of the DMRT132 gene
  cluster
• Final annotation of platypus DMRT gene
  cluster

• Expression of platypus DMRT genes

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Discussion

• Characteristics of platypus genome sequence
• Gene finding and annotating
• Location of DMRT1 genes on the sex chromosome
  chain
• Is DMRT1 involved in platypus sex determination?

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Thanks