Vatteroni L, Musio A, Meneveri R, Rainaldi G
Istituto di Mutagenesi e Differenziamento, Pisa, Italy.
Cytogenet Cell Genet. 1997;77(3-4):228-31. doi: 10.1159/000134582.
The Chinese hamster tumor-derived cell line 835T2 exhibits specific karyotypic changes, including the loss and/or translocation of genetic material. To investigate whether the p53 tumor suppressor gene was involved in the exchanges, cDNA from primary Chinese hamster cells was isolated by using sense and antisense primers of the human p53 gene. The cDNA was sequenced, and the sequence was compared with the Syrian and human p53 cDNA reported sequences. The sequence homology was very elevated, demonstrating that the cloned fragment contained part of the Chinese hamster p53 gene. The corresponding genomic fragment was also cloned and used as a biotin-labeled probe for in situ hybridization on Chinese hamster chromosome spreads. Hybridization was visualized by avidin-FITC, and the assignment was done comparing the banding obtained with BamHI restriction enzyme and the location of the fluorescent signals pattern of the same metaphase. The signals revealed that the p53 gene (TP53) is localized on Chinese hamster chromosome band 2p31, which is not involved in the karyotypic changes specific to the 835T2 cell line.
中国仓鼠肿瘤衍生细胞系835T2表现出特定的核型变化,包括遗传物质的丢失和/或易位。为了研究p53肿瘤抑制基因是否参与了这些交换,使用人p53基因的正义和反义引物从原代中国仓鼠细胞中分离出cDNA。对该cDNA进行测序,并将序列与已报道的叙利亚仓鼠和人p53 cDNA序列进行比较。序列同源性非常高,表明克隆片段包含中国仓鼠p53基因的一部分。相应的基因组片段也被克隆,并用作生物素标记探针,用于在中国仓鼠染色体涂片上进行原位杂交。通过抗生物素蛋白-异硫氰酸荧光素(avidin-FITC)观察杂交情况,并通过比较用BamHI限制酶获得的带型和同一中期荧光信号模式的位置来进行定位。信号显示p53基因(TP53)定位于中国仓鼠染色体2p31带,该带不参与835T2细胞系特有的核型变化。