Cytogenetic map of <i>Gekko hokouensis</i>, which shows chromosome homologies with the chicken and five squamate reptiles.

<p>This map was constructed with 86 functional genes and 18S–28S and 5S rRNA genes. Chromosomal locations of <i>ATP5A1</i>, <i>GHR</i>, <i>CHD1</i>, <i>DMRT1</i>, <i>RPS6</i>, and <i>ACO1</i>/<i>IREBP</i> were obtained from Kawai et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref039" target="_blank">39</a>]. The idiogram of <i>G</i>. <i>hokouensis</i> chromosomes was constructed according to Hoechst 33258-stained band patterns. Locations of the genes on <i>G</i>. <i>hokouensis</i> chromosomes are shown to the right of the chromosomes. The chromosome numbers show the chromosomes of the chicken (<i>Gallus gallus</i>, GGA), green anole (<i>Anolis carolinensis</i>, ACA), butterfly lizard (<i>Leiolepis reevesii rubritaeniata</i>, LRE), Japanese four-striped rat snake (<i>Elaphe quadrivirgata</i>, EQU), water monitor lizard (<i>Varanus salvator macromaculatus</i>, VSA), and sand lizard (<i>Lacerta agilis</i>, LAG), which show homologies with <i>G</i>. <i>hokouensis</i> chromosomes. no, no data on chromosome homology; un, a gene whose chromosomal location remains undetermined. Orange highlight indicates the genes that are homologous to chromosome segments of LAG. These genes are located on LRE, EQU, or VSA microchromosomes. The chromosomal locations of genes in the squamate reptiles were obtained from the following sources: <i>L</i>. <i>reevesii rubritaeniata</i> from Srikulnath et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref011" target="_blank">11</a>], <i>A</i>. <i>carolinensis</i> from Alföldi et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref014" target="_blank">14</a>], <i>E</i>. <i>quadrivirgata</i> from Matsubara et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref009" target="_blank">9</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref010" target="_blank">10</a>], <i>V</i>. <i>salvator macromaculatus</i> from Srikulnath et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref012" target="_blank">12</a>], and <i>L</i>. <i>agilis</i> from Srikulnath et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref013" target="_blank">13</a>].</p

Schematic representation for the process of chromosomal rearrangements that occurred among <i>Gekko hokouensis</i> chromosomes (GHO) 3, 6–8, and 12–15 and <i>Leiolepis reevesii rubritaeniata</i> chromosomes (LRE) 3–6.

<p>The diagram schematically summarizes the chromosomal rearrangements that occurred in LRE3, GHO13, and GHO15 (<b>a</b>); LRE4, GHO3, and GHO6 (<b>b</b>); LRE6, GHO8, and GHO12 (<b>c</b>); and LRE5, GHO7, and GHO14 (<b>d</b>). GHO6, GHO7, GHO12, and GHO15 are inverted to facilitate comparison. Chromosomal locations of the genes are shown to the right of the chromosomes by using arrowheads. Homologous chromosomes and/or chromosome segments are shown using the same color. Arrows indicate the directions of the chromosomal rearrangements.</p

List of 86 cDNA clones mapped to the Hokou gecko (<i>Gekko hokouensis</i>) chromosomes and their chromosomal locations in the sand lizard (<i>Lacerta agilis</i>), the water monitor lizard (<i>Varanus salvator macromaculatus</i>), the butterfly lizard (<i>Leiolepis reevesii rubritaeniata</i>), the Japanese four-striped rat snake (<i>Elaphe quadrivirgata</i>), the green anole (<i>Anolis carolinensis</i>), and the chicken (<i>Gallus gallus</i>).

<p>^aNucleotide sequences of two accession numbers were determined separately by forward and reverse primers in one clone.</p><p>^bThe cDNA fragment were obtain from <i>G</i>. <i>hokouensis</i>, which were mapped in our previous study (Kawai et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref039" target="_blank">39</a>]). For mapping of <i>ATP5A1</i>, <i>ACO1/IREBP</i> and <i>CHD1</i>, total length of cDNA fragment concatenated with multiple</p><p>–: No data</p><p>List of 86 cDNA clones mapped to the Hokou gecko (<i>Gekko hokouensis</i>) chromosomes and their chromosomal locations in the sand lizard (<i>Lacerta agilis</i>), the water monitor lizard (<i>Varanus salvator macromaculatus</i>), the butterfly lizard (<i>Leiolepis reevesii rubritaeniata</i>), the Japanese four-striped rat snake (<i>Elaphe quadrivirgata</i>), the green anole (<i>Anolis carolinensis</i>), and the chicken (<i>Gallus gallus</i>).</p

Chromosomal locations of the 18S–28S and 5S rRNA genes and (TTAGGG)n sequences in female <i>Gekko hokouensis</i>.

<p>(<b>a</b>) Hybridization pattern of FITC-labeled 18S–28S rRNA genes (green) and rhodamine-labeled 5S rRNA genes (red). Arrows indicate FISH signals of the 18S–28S rRNA genes, and arrowheads indicate signals of the 5S rRNA genes. (<b>b</b>) Hybridization pattern of FITC-labeled TTAGGG repeats (green) and rhodamine-labeled 5S rRNA genes (red). Arrows indicate signals of interstitial telomeric sites, and arrowheads indicate signals of the 5S rRNA genes. Scale bars represent 10 μm.</p

Comparative cytogenetic maps of macrochromosomes among <i>Gekko hokouensis</i>, <i>Lacerta agilis</i>, <i>Varanus salvator macromaculatus</i>, <i>Leiolepis reevesii rubritaeniata</i>, and <i>Elaphe quadrivirgata</i>, which were constructed with 68 functional genes.

<p>The chromosome map of <i>L</i>. <i>reevesii rubritaeniata</i> (LRE) was obtained from Srikulnath et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref011" target="_blank">11</a>]. The idiogram of <i>E</i>. <i>quadrivirgata</i> (EQU) macrochromosomes was obtained from Matsuda et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref008" target="_blank">8</a>] and chromosomal locations of the genes in <i>E</i>. <i>quadrivirgata</i>, from Matsubara et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref009" target="_blank">9</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref010" target="_blank">10</a>]. The chromosome map of <i>V</i>. <i>salvator macromaculatus</i> (VSA) and <i>L</i>. <i>agilis</i> (LAG) were obtained from Srikulnath et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref012" target="_blank">12</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0134829#pone.0134829.ref013" target="_blank">13</a>]. <i>G</i>. <i>hokouensis</i> chromosomes GHOZ, GHO6, GHO7, GHO12, and GHO15 and LAGZ, LAG8, LAG10, VSA3, VSA6, VSA7, EQU5, and EQU7 are inverted to facilitate comparison.</p

Hoechst 33258-stained karyotype of female <i>Gekko hokouensis</i>.

<p>Scale bar represents 10 μm.</p

Chromosomal locations of cDNA fragments of functional genes in female <i>Gekko hokouensis</i>.

<p><i>RBM12</i> was localized to chromosome 3 (GHO3) (<b>a</b>), <i>ATP2A2</i> to GHO11 (<b>c</b>), <i>SBNO1</i> to GHO11 (<b>d</b>), <i>SOX5</i> to GHO14 (<b>e</b>), <i>TLOC1</i> to GHO15 (<b>f</b>), <i>TMEM57</i> to GHO5 (<b>g</b>), and <i>WAC</i> to GHO8 (<b>h</b>). (<b>b, I, j, k, l, m, and n</b>) Hoechst 33258-stained patterns of the PI-stained metaphase spreads are shown in (<b>a, c, d, e, f, g,</b> and <b>h</b>). Arrows indicate the hybridization signals. Scale bars indicate 10 μm for (<b>a, b</b>) and 5 μm for (<b>c</b>–<b>h</b>).</p

Table_S5

Frequencies of nonsynonymous/synonymous substitution ratios (Ka/Ks) in exon 11 of the CTNNB1 gene among eight snake species. The CTNNB1W gene is shown above the diagonal with the CTNNB1Z gene below the diagonal. Table is also available in PDF format in the "Supplementary file" PD

Supplementary file

Supplementary file includes supplementary figures and tables, which are relevant to publication. Please note that the supplementary tables included in this PDF are also uploaded separately, as .xlsx files

Table_S2

Pair-wise comparison of amino acid sequence identities (%) of CTNNB1W among eight snake species. Table is also available in PDF format in the "Supplementary file" PD