Kuroiwa Y, Tomizuka K, Shinohara T, Kazuki Y, Yoshida H, Ohguma A, Yamamoto T, Tanaka S, Oshimura M, Ishida I
Pharmaceutical Research Laboratory, Kirin Brewery. Co., Ltd., 3 Miyahara-cho Takasaki-shi Gunma 370-1295, Japan.
Nat Biotechnol. 2000 Oct;18(10):1086-90. doi: 10.1038/80287.
For introducing regions of human chromosomes greater than a megabase into cells or animals, we have developed a chromosome-cloning system in which defined regions of human chromosomes can be cloned into a stable human minichromosome vector in homologous recombination-proficient chicken DT40 cells. The stable minichromosome vector allowed a 10 Mb-sized region of the mitotically unstable human chromosome 22 to be stably maintained in mouse embryonic stem (ES) cells, and in mice. Furthermore, we demonstrated functional expression of human genes from the HAC in mice. This study describes a stable cloning and expression system for greater than megabase-sized regions of human chromosomes.
为了将大于1兆碱基的人类染色体区域导入细胞或动物体内，我们开发了一种染色体克隆系统，在该系统中，人类染色体的特定区域可在同源重组能力强的鸡DT40细胞中克隆到稳定的人类微型染色体载体中。这种稳定的微型染色体载体使有丝分裂不稳定的人类22号染色体上一个10兆碱基大小的区域能在小鼠胚胎干细胞和小鼠体内稳定维持。此外，我们还证明了小鼠体内源自人人工染色体（HAC）的人类基因的功能性表达。本研究描述了一种用于人类染色体大于兆碱基大小区域的稳定克隆和表达系统。
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