Cloning a fragment from the telomere of the long arm of human chromosome 9 in a YAC vector.
The construction of a yeast artificial chromosome containing a human DNA insert is reported. This molecule of about 200 kb behaves as a native yeast chromosome since it has a very high mitotic stability and is present in the yeast transformant clone at a copy number similar to that of the resident chromosomes. Hybridization with the TTAGGG sequence demonstrates that this chromosome contains human telomeric sequences. In situ hybridization of the biotin-labelled artificial chromosome to metaphase human chromosomes shows that the insert occupies a telomeric position on the long arm of chromosome 9. Since the fragment was cloned as an EcoRI insert and not as a telomere, it is situated medially to the telomeric sequences and harbours telomere-associated sequences, that have been shown to contain the TTAGGG sequence. The fragment represents the end of the genetic map of chromosome 9 and thus can be used to characterize the sequence and the structure of the chromosomal region that runs from the end of the chromosome to the first gene.
Guerrini
A M
AM
Dipartimento di Biologia Cellulare e dello Sviluppo, Universitá La Sapienza, Roma, Italy.
Ascenzioni
F
F
Pisani
G
G
Rappazzo
G
G
Della Valle
G
G
Donini
P
P
D001483:Base Sequence; D015139:Blotting, Southern; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D002899:Chromosomes, Human, Pair 9; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic
D004251:DNA Transposable Elements