Sun T Q, Fenstermacher D A, Vos J M
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill 27599.
Nat Genet. 1994 Sep;8(1):33-41. doi: 10.1038/ng0994-33.
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.
我们基于大型疱疹病毒爱泼斯坦-巴尔病毒的潜在复制起点，开发了一种人类人工附加体染色体（HAEC）系统，用于在人类细胞中将DNA作为环状微型染色体进行扩增和稳定维持。单个HAEC携带60 - 330 kb的人类基因组插入片段，并且在遗传上表现稳定。我们建立了一个包含1500个独立克隆的HAEC文库，这些克隆携带平均大小为150 - 200 kb的随机人类基因组片段，并能够回收HAEC DNA。我们的自体HAEC系统将人类DNA直接克隆到人类细胞中，为大型哺乳动物DNA区域的功能研究和基因治疗提供了一个重要工具。
