Construction of YAC-based mammalian artificial chromosomes.
To construct a mammalian artificial chromosome (MAC), telomere repeats and selectable markers were introduced into a 100 kb yeast artificial chromosome (YAC) containing human centromeric DNA. This YAC, which has a regular repeat structure of alpha-satellite DNA and centromere protein B (CENP-B) boxes, efficiently formed MACs that segregated accurately and bound CENP-B, CENP-C, and CENP-E. The MACs appear to be about 1-5 Mb in size and contain YAC multimers. Structural analyses suggest that the MACs have not acquired host sequences and were formed by a de novo mechanism. The accurate segregation of the MACs suggests they have potential as vectors for introducing genes into mammals.
Ikeno
M
M
Department of Molecular Biology, School of Science, Nagoya University, Japan.
Grimes
B
B
Okazaki
T
T
Nakano
M
M
Saitoh
K
K
Hoshino
H
H
McGill
N I
NI
Cooke
H
H
Masumoto
H
H
D000818:Animals; D001324:Autoantigens; D002460:Cell Line; D002461:Cell Line, Transformed; D002503:Centromere / Q000235:genetics; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics; D002875:Chromosomes / Q000235:genetics*; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics; D003001:Cloning, Molecular; D004276:DNA, Satellite / Q000737:chemistry / Q000235:genetics; D004268:DNA-Binding Proteins; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018386:Kinetochores; D016133:Polymerase Chain Reaction; D016615:Telomere / Q000235:genetics; D014162:Transfection
D001324:Autoantigens; C497926:CENPB protein, human; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D004276:DNA, Satellite; D004268:DNA-Binding Proteins; C060905:centromere protein C; C068688:centromere protein E