Human mini-chromosomes with minimal centromeres.
We have introduced a 6.5 Mb human mini-chromosome with a complex centromere structure into DT40 cells and have used sequence targeting and telomere-directed chromosome breakage to dissect the sequence requirements for centromere function. These experiments proved that a vertebrate centromere with two blocks of functional alphoid DNA separated by 2.5 Mb can exist as a stable structure in some but not all vertebrate cells. Further experiments indicated that recovery of chromosomes with less than approximately 100 kb of alphoid DNA is very inefficient, suggesting that a functional centromere requires a minimum of approximately 100 kb of alphoid DNA. Mini-chromosomes with minimal centromeres segregate accurately in some but not all vertebrate cells and should be useful for the detection of sequence-specific factors required for vertebrate centromere maintenance.
Biochemistry Department, South Parks Road, Oxford OX1 3QU, UK.
J W
JW
Yang
C
C
Pendon
J
J
Yang
N
N
Haywood
A
A
Chand
W R
WR
Brown
D000818:Animals; D002460:Cell Line; D002503:Centromere; D002645:Chickens; D002877:Chromosomes, Human; D006801:Humans