A novel method of caenophidian snake sex identification using molecular markers based on two gametologous genes |
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| Authors: | Nararat Laopichienpong Panupong Tawichasri Lawan Chanhome Rattanin Phatcharakullawarawat Worapong Singchat Attachai Kantachumpoo Narongrit Muangmai Sunutcha Suntrarachun Kazumi Matsubara Surin Peyachoknagul Kornsorn Srikulnath |
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| Affiliation: | 1. Laboratory of Animal Cytogenetics and Comparative Genomics (ACCG), Department of Genetics, Faculty of Science, Kasetsart University, Bangkok, Thailand;2. Animal Breeding and Genetics Consortium of Kasetsart University (ABG ‐ KU), Bangkok, Thailand;3. Snake Farm, Queen Saovabha Memorial Institute, The Thai Red Cross Society, Bangkok, Thailand;4. Mildpets Animal Hospital, Bangkok, Thailand;5. Center for Advanced Studies in Tropical Natural Resources, National Research University‐Kasetsart University, Thailand (CASTNAR, NRU‐KU, Thailand), Kasetsart University, Bangkok, Thailand;6. Department of Fishery Biology, Faculty of Fisheries, Kasetsart University, Bangkok, Thailand;7. Department of Research and Development, Queen Saovabha Memorial Institute, The Thai Red Cross Society, Bangkok, Thailand;8. Research Center for Bioinformatics and Biosciences, National Research Institute of Fisheries Science, Japan Fisheries Research and Education Agency, Yokohama, Kanagawa, Japan;9. Department of Biology, Faculty of Science, Naresuan University, Phitsanulok, Thailand |
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| Abstract: | Sex identification provides important information for ecological and evolutionary studies, as well as benefiting snake conservation management. Traditional methods such as cloacal probing or cloacal popping are counterproductive for sex identification concerning very small species, resulting in difficulties in the management of their breeding programs. In this study, the nucleotide sequences of gametologous genes (CTNNB1 and WAC genes) were used for the development of molecular sexing markers in caenophidian snakes. Two candidate markers were developed with the two primer sets, and successfully amplified by a single band on the agarose gel in male (ZZ) and two bands, differing in fragment sizes, in female (ZW) of 16 caenophidian snakes for CTNNB1 and 12 caenophidian snakes for WAC. Another candidate marker was developed with the primer set to amplify the specific sequence for CTNNB1W homolog, and the PCR products were successfully obtained in a female‐specific 250‐bp DNA bands. The three candidate PCR sexing markers provide a simple sex identification method based on the amplification of gametologous genes, and they can be used to facilitate effective caenophidian snake conservation and management programs. |
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| Keywords: |
CTNNB1
gametolog sex chromosome snake WAC |
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