Isolation and identification of a novel tandemly repeated DNA sequence in the centromeric region of human chromosome 8 |
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| Authors: | C C Lin R Sasi Y S Fan D Court |
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| Institution: | (1) Department of Laboratory Medicine and Pathology, University of Alberta, T6G2B7 Edmonton, Alberta, Canada;(2) University of Alberta Hospitals, T6G 2B7 Edmonton, Alberta, Canada;(3) Present address: Cytogenetics Laboratory, Department of Laboratory Medicine, Credit Valley Hospital, 2200 Eglinton Avenue West, L5M 2N1 Mississauga, Ontario, Canada;(4) Cytogenetics Laboratory, 5B4,49 W.C. Mackenzie Health Sciences Centre, University of Alberta Hospitals, 8440-112 Street, T6G 2B7 Edmonton, Alberta, Canada |
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| Abstract: | EcoRI subclones, designated as 50E1 and 50E4, were independently obtained from a cosmid clone previously mapped to the centromeric region of human chromosome 8. Southern blot hybridization analyses suggested that both subclones contain repetitive DNA sequences different from the chromosome 8 specific alphoid DNA. DNA sequence analysis of the 704 bp insert of 50E1 and the 1, 962 bp insert of 50E4 revealed that both inserts contained tandemly repeated units of  220 bp. Fluorescence in situ hybridization studies confirmed these two subclones to be specifically located on the centromeric region of chromosome 8. A 220 bp consensus sequence, derived from nine monomeric repeats, showed no significant homology to alphoid consensus sequences or to other currently known human centromeric DNA sequence. Furthermore, no significant homology was found with any other DNA sequence deposited in the EMBL or GenBank databases, indicating that this chromosome 8 specific repetitive DNA sequence is novel. From slot blot experiments it was estimated that 0.013% of the human genome comprises 1,750 of these monomeric repeats, residing on the centromeric region of chromosome 8 in tandem array(s). |
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