Abstract
A minilibrary was constructed from DOP-PCR products using microdissected Y-chromosomes of Indian muntjac as DNA templates. Two microclones designated as IM-Y4-52 and IM-Y5-7 were obtained from negative screening of all three cervid satellite DNAs (satellites I, II, and IV). These two microclones were 295 and 382 bp in size, respectively, and shared ∼70% sequence homology. Southern blot analysis showed that the IM-Y4-52 clone was repetitive in nature with an ∼0.32-kb register in HaeIII digest. Sequence comparison revealed no similarities to DNA sequences deposited in the GenBank database, suggesting that the microclone sequences were from a novel satellite DNA family designated as cervid satellite V. A subclone of an Indian muntjac BAC clone which screened positive for IM-Y4-52 had a 3,325-bp insert containing six intact monomers, four deleted monomers, and two partial monomers. The consensus sequence of the monomer was 328 bp in length and shared more than 80% sequence homology with every intact monomer. A zoo blot study using IM-Y4-52 as a probe showed that the strong hybridization with EcoRI digested male genomic DNA of Indian muntjac, Formosan muntjac, Chinese muntjac, sambar deer, and Chinese water deer. Female genomic DNA of Indian muntjac, Chinese water deer, and Formosan muntjac also showed positive hybridization patterns. Satellite V was found to specifically localize to the Y heterochromatin region of the muntjacs, sambar deer, and Chinese water deer and to chromosome 3 of Indian muntjac and the X-chromosome of Chinese water deer.
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Acknowledgements
This study was supported by grants from the National Science Council, Taiwan (NSC92-2320-B-040-048), and from the National Health Research Institute, Taiwan (NHRI-EX92-9207SI). We thank Leona Chemnick and Susan Hansen for help with sample inventory and transfer, including documentation required under US regulations.
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Y.-C. Li and Y.-M. Cheng contributed equally to this work.
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Li, YC., Cheng, YM., Hsieh, LJ. et al. Karyotypic evolution of a novel cervid satellite DNA family isolated by microdissection from the Indian muntjac Y-chromosome. Chromosoma 114, 28–38 (2005). https://doi.org/10.1007/s00412-005-0335-7
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DOI: https://doi.org/10.1007/s00412-005-0335-7