HUMAN ARTIFICIAL EPISOMAL CHROMOSOMES FOR CLONING LARGE DNA FRAGMENTS IN HUMAN-CELLS

被引:70
作者
SUN, TQ
FENSTERMACHER, DA
VOS, JMH
机构
[1] UNIV N CAROLINA, DEPT BIOCHEM & BIOPHYS, CHAPEL HILL, NC 27599 USA
[2] UNIV N CAROLINA, CURRICULUM GENET & MOLEC BIOL, CHAPEL HILL, NC 27599 USA
[3] UNIV N CAROLINA, LINEBERGER COMPREHENS CANC CTR, CHAPEL HILL, NC 27599 USA
关键词
D O I
10.1038/ng0994-33
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
We have developed a human artificial episomal chromosome (HAEC) system, based on the latent replication origin of the large herpes Epstein-Barr virus, for the propagation and stable maintenance of DNA as circular minichromosomes in human cells. Individual HAECs carried human genomic inserts ranging from 60-330 kb and appeared genetically stable. An HAEC library of 1,500 independent clones carrying random human genomic fragments with average sizes of 150-200 kb was established and allowed recovery of the HAEC DNA. Our autologous HAEC system, with human DNA cloned directly in human cells, provides an important tool for functional study of large mammalian DNA regions and gene therapy.
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页码:33 / 41
页数:9
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