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为了探讨p16基因在人肿瘤细胞系中的突变,采用PCR-SSCP分析和基因序列测定技术,对人黑色素瘤细胞系(Bowes)中p16基因的突变进行检测。结果显示,Bowes细胞的p16基因第二外显子扩增产物在聚丙烯酰胺凝胶中电泳异常,表明其基因序列存在点突变。核酸序列测定证实第341号核苷酸发生T→C的颠换。应用DNA重组技术构建的突变型p16基因重组质粒可作为基因探针用于检测p16基因的缺失。
In order to investigate the mutation of p16 gene in human tumor cell lines, mutations of p16 gene in human melanoma cell lines (Bowes) were detected by PCR-SSCP analysis and gene sequencing. The results showed that the amplified product of the second exon of the p16 gene in Bowes cells was abnormally electrophoresed on a polyacrylamide gel, indicating that there was a point mutation in the gene sequence. Nucleic acid sequencing confirmed the transversion of T→C in nucleotide 341. Mutant p16 gene recombinant plasmid constructed by using DNA recombination technology can be used as a gene probe to detect deletion of p16 gene.