Karyotypic evolution of a novel cervid satellite DNA family isolated by microdissection from the Indian muntjac Y-chromosome |
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Authors: | Y.-C. Li Y.-M. Cheng L.-J. Hsieh O. A. Ryder F. Yang S.-J. Liao K.-M. Hsiao F.-J. Tsai C.-H. Tsai C. C. Lin |
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Affiliation: | (1) Department of Biomedical Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien Kuo N. Rd., 40203 Taichung, Taiwan;(2) Department of Medical Research, China Medical University and Hospital, No. 2, Yuh-Der Road, 404 Taichung, Taiwan;(3) Center for Reproduction of Endangered Species, Zoological Society of San Diego, PO Box 120551, San Diego, CA 92112-0551, USA;(4) Center for Veterinary Science, University of Cambridge, Madingley Road, Cambridge, CB3 0ES, UK |
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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.Y.-C. Li and Y.-M. Cheng contributed equally to this work. |
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