Efficiency of de novo centromere formation in human artificial chromosomes

被引：64

作者：

Mejía, JE

Alazami, A

Willmott, A

Marschall, P

Levy, E

Earnshaw, WC

Larin, Z ^{[1
]}

机构：

[1] Univ Oxford, Weatherall Inst Mol Med, Oxford OX3 7BN, England

[2] Univ Oxford, Wellcome Trust Ctr Human Genet, Oxford OX3 7BN, England

[3] Univ Edinburgh, Inst Cell & Mol Biol, Edinburgh EH9 3JR, Midlothian, Scotland

来源：

GENOMICS | 2002年 / 79卷 / 03期

基金：

英国惠康基金;

关键词：

centromere; chromosomes; artificial chromosomes; human; pair; 17; DNA; satellite; Y chromosome;

D O I：

10.1006/geno.2002.6704

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

In a comparative study, we show that human artificial chromosome (HAC) vectors based on ut-satellite (alphoid) DNA from chromosome 17 but not the Y chromosome regularly form HACs in HT1080 human cells., We constructed four structurally similar HAC vectors, two with chromosome 17 or Y alphoid DNA (17alpha, Yalpha) and two with 17a or Ya and the hypoxanthine guanine phosphoribosyltransferase locus (HPRT1). The 17alpha HAC vectors generated artificial minichromosomes in 32-79% of the HT1080 clones screened, compared with only similar to 4% for the Yet HAC vectors, indicating that Ya is inefficient at forming a de novo centromere. The 17alpha HAC vectors produced megabase-sized, circular HACs containing multiple copies of alphoid fragments (60-250 kb) interspersed with either vector or HPRT1 DNA. The 17alpha-HPRT1 HACs were less stable than those with 17alpha only, and these results may influence the design of new HAC gene transfer vectors.

引用

页码：297 / 304

页数：8

共 36 条

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