Construction of BAC-based physical map and analysis of chromosome rearrangement in chinese hamster ovary cell lines
Construction of BAC-based physical map and analysis of chromosome rearrangement in chinese hamster ovary cell lines
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DOI：
10.1002/bit.24347
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发表时间：
2012-06-01
影响因子：
3.8
通讯作者：
Omasa, Takeshi
中科院分区：
文献类型：
--
作者：
Cao, Yihua;Kimura, Shuichi;Omasa, Takeshi
中国仓鼠卵巢细胞（CHO）作为哺乳动物宿主细胞克隆和表达相关基因的平台，多年来一直被广泛应用于生物技术。随机选择303个BAC克隆作为杂交探针（BAC-FISH），采用荧光原位杂交（FISH）技术构建CHO DG44细胞系的详细物理染色体图谱。最长的两条染色体是完全成对的；其他染色体部分被删除或重新排列。对624个BAC克隆（其中287个已定位BAC克隆）的末端序列进行分析，得到1119个信息性BAC末端序列。303个BAC克隆中，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的BAC- fish分析。基于所构建的物理图谱和末端序列，研究CHO DG44、CHO K1与中国仓鼠原代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保守，未发生大的重排。这一结果表明，这些染色体在CHO细胞中是稳定和必需的，并且在其他CHO细胞系中可能是保守的。Biotechnol。Bioeng。2012;109:13571367。(c) 2011 Wiley期刊公司
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. Biotechnol. Bioeng. 2012; 109:13571367. (c) 2011 Wiley Periodicals, Inc.