Molecular cloning of human telomeres in yeast.
Telomeres are the DNA sequences found at the ends of linear chromosomes. They define the boundaries of the genetical and physical maps of such chromosomes and so are particularly important for the complete mapping of large genomes that is now being attempted. Telomeres have been intensively studied in the yeast Saccharomyces cerevisiae and in ciliated protozoa: in these organisms the telomeric DNA consists of arrays of tandemly repeated short sequences in which one strand is guanosine-rich and oriented 5' to 3' towards the chromosome end. The conservation of these structural features is reflected in the observation that telomeric DNA from a variety of protozoa will function as telomeres on artificial linear mini-chromosomes in yeast. Tandem arrays of the sequence TTAGGG have been identified at the telomeres of humans and other mammals and also of trypanosomes. This indicates that the structural features of telomeres are conserved between higher and lower eukaryotes and implies that human telomeric DNA could function in yeast. I have used this idea to develop a strategy to isolate a specific human telomere as a molecular clone in yeast and have devised a simple and effective way of cloning other human telomeres and their associated sequences.
Brown
W R
WR
Biochemistry Department, Oxford University, UK.
D000818:Animals; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D002877:Chromosomes, Human / Q000648:ultrastructure*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; D004587:Electrophoresis, Agar Gel; D005822:Genetic Vectors; D006801:Humans; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic; D014345:Trypanosoma / Q000235:genetics
D015345:Oligonucleotide Probes; D004247:DNA; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI