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Open Access Research article

Testes-specific hemoglobins in Drosophila evolved by a combination of sub- and neofunctionalization after gene duplication

Eva Gleixner12, Holger Herlyn3, Stefan Zimmerling1, Thorsten Burmester4 and Thomas Hankeln1*

Author Affiliations

1 Institute of Molecular Genetics, University of Mainz, 55099 Mainz, Germany

2 Center for Systems Biology, University of Freiburg, 79104 Freiburg, Germany

3 Institute of Anthropology, University of Mainz, 55099 Mainz, Germany

4 Biocenter Grindel and Zoological Museum, University of Hamburg, 20146 Hamburg, Germany

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BMC Evolutionary Biology 2012, 12:34  doi:10.1186/1471-2148-12-34

Published: 19 March 2012

Additional files

Additional file 1:

Designations of Drosophila globin genes.

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Additional file 2:

Phylogenetic relationship of Drosophila globins. Phylogenetic reconstruction of Drosophila glob1, glob2, glob3 and glob2/3 including G. intestinalis glob1 (ginglob1) and C. thummi thummi HbIII (cttHbIII) at the nucleotide level (using only first and second codon positions). (A) by applying a Maximum likelihood approach implemented in Treefinder and (B) a Bayesian analysis using MrBayes.

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Additional file 3:

Absolute mRNA quantitation of dmeglob2, dmeglob3 and dviglob2/3. Glob2, glob3 and glob2/3 absolute mRNA copy number in male adult flies of D. melanogaster and D. virilis, measured with qPCR. Dmeglob2 (144784 copies) and dmeglob3 (150026 copies) copy number summed up is equal to dviglob2/3 copy number (311673 copies).

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Additional file 4:

Glob3 gene region with Jockey transposon. Schematic diagram of the genomic region of D. melanogaster glob3 including the transposable element inserted downstream of the glob3 gene in comparison to the corresponding genomic region of D. sechellia glob3. In D. melanogaster, the insertion and the putative duplicated sequences are indicated. In D. sechellia, the predicted promoter sequence spanning the transposon insertion sequence in D. melanogaster are highlighted. Exons and 5'UTR and distances between 3'end of transposable element and Exon1 in both D. melanogaster and D. sechellia are plotted.

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Additional file 5:

Negative control for mRNA in situ hybridization. As negative control for in situ hybridization, a sense mRNA probe of dmeglob2 was applied at otherwise identical hybridization conditions.

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Additional file 6:

Regulation of LDH mRNA in D. melanogaster male adults after hypoxic stress. mRNA levels (bars) are shown relative to the gene expression at normoxia (21%). (A) LDH expression after 1% O2 for 1 h, 3 h and 4 h. After 1 h of hypoxia, no alteration in LDH expression could be detected. 3 h of hypoxia caused the LDH mRNA levels to increase about 2.2 fold and 4 h of hypoxia to about 3.4 fold compared to the normoxic control (B) LDH expression after 6% O2for 24 h. After applying long-term moderate hypoxia with 6% O2 for 24 h, an increase in LDH mRNA expression to about 1.5 fold could be detected (*p < 0.05; **p < 0.01).

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Additional file 7:

Oligonucleotides and PCR conditions

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