Efficient construction of BAC-based human artificial chromosomes (HACs) requires optimization of each key functional unit as well as development of techniques for the rapid and reliable manipulation of high-molecular weight BAC vectors. Here, we have created synthetic chromosome 17-derived alpha-satellite arrays, based on the 16-monomer repeat length typical of natural D17Z1 arrays, in which the consensus CENP-B box elements are either completely absent (0/16 monomers) or increased in density (16/16 monomers) compared to D17Z1 alpha-satellite (5/16 monomers). Using these vectors, we show that the presence of CENP-B box elements is a requirement for efficient de novo centromere formation and that increasing the density of CENP-B box elements may enhance the efficiency of de novo centromere formation. Furthermore, we have developed a novel, high-throughput methodology that permits the rapid conversion of any genomic BAC target into a HAC vector by transposon-mediated modification with synthetic alpha-satellite arrays and other key functional units. Taken together, these approaches offer the potential to significantly advance the utility of BAC-based HACs for functional annotation of the genome and for applications in gene transfer.
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Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Ebersole, TA
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Ross, A
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Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Ross, A
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Clark, E
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McGill, N
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Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
McGill, N
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Schindelhauer, D
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Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Schindelhauer, D
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Cooke, H
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Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Cooke, H
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Grimes, B
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h-index: 0
机构:
Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
机构:
Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Ebersole, TA
;
Ross, A
论文数: 0引用数: 0
h-index: 0
机构:
Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Ross, A
;
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机构:
Clark, E
;
McGill, N
论文数: 0引用数: 0
h-index: 0
机构:
Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
McGill, N
;
Schindelhauer, D
论文数: 0引用数: 0
h-index: 0
机构:
Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Schindelhauer, D
;
Cooke, H
论文数: 0引用数: 0
h-index: 0
机构:
Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland
Cooke, H
;
Grimes, B
论文数: 0引用数: 0
h-index: 0
机构:
Univ Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, ScotlandUniv Edinburgh, Western Gen Hosp, MRC, Human Genet Unit, Edinburgh EH4 2XU, Midlothian, Scotland