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基于 BAC 的物理图谱构建和中国仓鼠卵巢细胞系染色体重排分析。

Construction of BAC-based physical map and analysis of chromosome rearrangement in Chinese hamster ovary cell lines.

机构信息

Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.

出版信息

Biotechnol Bioeng. 2012 Jun;109(6):1357-67. doi: 10.1002/bit.24347. Epub 2011 Nov 6.

Abstract

Chinese hamster ovary (CHO) cells have frequently been used in biotechnology for many years as a mammalian host cell platform for cloning and expressing genes of interest. A detailed physical chromosomal map of the CHO DG44 cell line was constructed by fluorescence in situ hybridization (FISH) imaging using randomly selected 303 BAC clones as hybridization probes (BAC-FISH). The two longest chromosomes were completely paired chromosomes; other chromosomes were partly deleted or rearranged. The end sequences of 624 BAC clones, including 287 mapped BAC clones, were analyzed and 1,119 informative BAC end sequences were obtained. Among 303 mapped BAC clones, 185 clones were used for BAC-FISH analysis of CHO K1 chromosomes and 94 clones for primary Chinese hamster lung cells. Based on this constructed physical map and end sequences, the chromosome rearrangements between CHO DG44, CHO K1, and primary Chinese hamster cells were investigated. Among 20 CHO chromosomes, eight were conserved without large rearrangement in CHO DG44, CHO K1, and primary Chinese hamster cells. This result suggested that these chromosomes were stable and essential in CHO cells and supposedly conserved in other CHO cell lines.

摘要

中国仓鼠卵巢(CHO)细胞多年来一直被广泛应用于生物技术领域,作为克隆和表达目的基因的哺乳动物宿主细胞平台。通过使用随机选择的 303 个 BAC 克隆作为杂交探针的荧光原位杂交(FISH)成像,构建了 CHO DG44 细胞系的详细物理染色体图谱(BAC-FISH)。两条最长的染色体完全配对;其他染色体部分缺失或重排。分析了 624 个 BAC 克隆的末端序列,包括 287 个已定位的 BAC 克隆,获得了 1119 个信息性 BAC 末端序列。在 303 个已定位的 BAC 克隆中,有 185 个克隆用于 CHO K1 染色体的 BAC-FISH 分析,94 个克隆用于原代中国仓鼠肺细胞。基于这个构建的物理图谱和末端序列,研究了 CHO DG44、CHO K1 和原代中国仓鼠细胞之间的染色体重排。在 20 条 CHO 染色体中,有 8 条在 CHO DG44、CHO K1 和原代中国仓鼠细胞中没有大的重排而被保守保留。这一结果表明,这些染色体在 CHO 细胞中是稳定和必需的,在其他 CHO 细胞系中可能也被保守保留。

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