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一种表达大鼠CYP2B1的基因工程V79细胞系SD1在转染DNA的整合位点表现出染色体不稳定性。

A genetically engineered V79 cell line SD1 expressing rat CYP2B1 exhibits chromosomal instability at the integration site of the transfected DNA.

作者信息

Ellard S, James S A, Parry E M, Parry J M

机构信息

School of Biological Sciences, University of Wales, Singleton Park, Swansea, UK.

出版信息

Mutagenesis. 1995 Nov;10(6):549-54. doi: 10.1093/mutage/10.6.549.

Abstract

The genetically engineered cell line SD1 was constructed by co-transfection of V79 Chinese hamster cells with two plasmids: one containing a full-length cDNA encoding rat CYP2B1 and the second incorporating a selective marker gene. This cell line has been used in gene mutation tests and in cytokinesis-block micronucleus assays to identify procarcinogens which are metabolized by CYP2B1 to reactive metabolites. An elevated frequency of spontaneous micronuclei was recorded in SD1 cells compared to parental V79 cultures. Karyotypic analysis revealed a chromosomal instability which was manifested by amplification of the p-arms of a chromosome designated 'n' (derived from chromosome 8). This chromosome was variable in length and sometimes exhibited a telomeric fusion which led to the formation of a dicentric chromosome. Fluorescence in situ hybridization with digoxigenin-labelled plasmid DNA showed the presence of pSV450 plasmid DNA coamplified with genomic DNA sequences located in the terminal region of chromosome 'n'.

摘要

通过将V79中国仓鼠细胞与两种质粒共转染构建了基因工程细胞系SD1:一种质粒含有编码大鼠CYP2B1的全长cDNA,另一种质粒含有选择性标记基因。该细胞系已用于基因突变试验和胞质分裂阻断微核试验,以鉴定被CYP2B1代谢为活性代谢物的前致癌物。与亲代V79培养物相比,SD1细胞中自发微核的频率升高。核型分析显示存在染色体不稳定性,表现为一条命名为“n”的染色体(源自8号染色体)的p臂扩增。这条染色体长度可变,有时会出现端粒融合,导致双着丝粒染色体的形成。用地高辛标记的质粒DNA进行荧光原位杂交显示,pSV450质粒DNA与位于染色体“n”末端区域的基因组DNA序列共同扩增。

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