Increase of homozygosity in centenarians revealed by a new inter-Alu PCR technique.
In the present study a novel inter-Alu PCR technique that allows one to detect inter-individual differences in the genomic regions flanked by Alu repetitive sequences was developed. Two primers complementary to sequences present in different Alu repeats and marked with two different fluorochromes were used in the same PCR reaction, and the PCR products were separated and analyzed by capillary electrophoresis using an automatic sequencer. The method is highly reliable, and three patterns of peaks (QM376-400, QM780-790 and QM480) appeared to be representative for germ-line polymorphisms, as suggested by the results obtained in nine couples of monozygotic twins and four three-generation families. The frequency of these polymorphic peaks was studied in two different age groups (100 young subjects and 69 centenarians). In two out of the three regions (QM376-400 and QM480) a significant increase in homozygote genotypes frequency was observed in centenarians. These counterintuitive results suggest that increased homozygosity contributes to human longevity. This novel inter-Alu PCR approach could represent a valuable tool to identify longevity-associated DNA sequences interspersed throughout human genome, without making any a priori assumption about their nature and function.
Bonafè
M
M
Department of Experimental Pathology, University of Bologna, Bologna, Italy.
Cardelli
M
M
Marchegiani
F
F
Cavallone
L
L
Giovagnetti
S
S
Olivieri
F
F
Lisa
R
R
Pieri
C
C
Franceschi
C
C
D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000235:genetics*; D020087:Alu Elements; D006579:Heterozygote; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic