Separation of long-range human TERT gene haplotypes by transformation-associated recombination cloning in yeast.
The hTERT gene encoding a catalytic subunit of human telomerase contains four blocks of variable number of tandem repeats (VNTRs)--two in intron 2 and two in intron 6. The segregation of hTERT VNTRs was analysed in families, revealing that all of them were transmitted through meiosis following a Mendelian inheritance. The work reports a further characterization of the minisatellites in hTERT. We employed transformation-associated recombination (TAR) cloning to isolate parental hTERT alleles and determined the specific combination of minisatellites at each of the polymorphic sites. A long-range haplotyping of hTERT determined by TAR cloning was verified by classical Mendelian analysis. Since such a strategy can be applied for any chromosomal locus, we conclude that recombinational gene capture could greatly facilitate haplotypes analysis.
Kim
Jung-Hyun
JH
Laboratory of Biosystems and Cancer, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Leem
Sun-Hee
SH
Sunwoo
Yangil
Y
Kouprina
Natalay
N
D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D005260:Female; D006239:Haplotypes; D006801:Humans; D008297:Male; D018598:Minisatellite Repeats; D011110:Polymorphism, Genetic; D019098:Telomerase / Q000235:genetics*; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
D004268:DNA-Binding Proteins; C509186:TERT protein, human; D019098:Telomerase