A non-alphoid repetitive DNA sequence from human chromosome 21.
A non-alphoid repetitive DNA from human chromosome 22, consisting of a 48-bp motif, shows homology to both G-group chromosomes in the gorilla, thus indicating the presence of additional repeat family members on further human chromosomes. Therefore, we screened a chromosome-21-specific cosmid library using this repetitive sequence from chromosome 22 (D22Z3). Some 40-50 cosmid clones were positive in tests for hybridization. One of the clones giving the strongest signals was digested with EcoRI/PstI, which we knew to cut frequently within the repeats; this resulted in fragments containing repeat units only. The fragments were subcloned into plasmid vector pTZ 19. Sequence-analysis of a 500-bp insert showed ten copies of a 48-bp repeat similar to D22Z3, with about 15% sequence deviation from the chromosome 22 consensus sequence. In situ hybridization of the newly isolated recombinant established its chromosome 21 specificity at high stringency. Physical mapping by pulsed field gel electrophoresis placed this new repeat in close vicinity to the chromosome 21 alphoid repeat. No cross-hybridization with other mammalian genomes except for those of apes was observed. The locus has been designated D21Z2 by the Genome Data Base. A gel mobility shift assay indicated that this repetitive motif has protein-binding properties.
Müllenbach
R
R
Institut für Humangenetik, Universität des Saarlandes, Homburg/Saar, Federal Republic of Germany.
Lutz
S
S
Holzmann
K
K
Dooley
S
S
Blin
N
N
D000818:Animals; D001483:Base Sequence; D002503:Centromere / Q000737:chemistry; D002869:Chromosome Aberrations; D002891:Chromosomes, Human, Pair 21; D003001:Cloning, Molecular; D003360:Cosmids; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004285:Dogs; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011323:Primates; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D015183:Restriction Mapping; D012377:Rodentia; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013552:Swine
D015342:DNA Probes; D004247:DNA