U87胶质瘤细胞p53基因全场cDNA克隆及序列分析

Cloning and analysising of the full-length cDNA of p53 gene in U87 glioma cells

目的克隆U87胶质瘤细胞p53基因编码序列,并对其序列进行分析。方法根据NCBI GenBank中p53 cDNA序列设计引物,利用RT-PCR技术从U87胶质瘤细胞总RNA中对p53基因编码序列进行克隆;并通过DNAMAN软件将克隆序列与NCBI GenBank中序列进行比对,分析U87细胞中p53基因编码序列特点。结果经测序验证,成功克隆了p53基因编码序列,通过序列比对发现U87细胞中p53基因72位密码子为精氨酸残基。结论成功克隆了U87细胞中p53基因编码序列,分析发现U87细胞中p53基因表达产生的是72位密码子为含精氨酸残基的p53野生型蛋白,为进一步研究U87细胞中p53基因功能特性及其与胶质瘤的相关性奠定了基础。

Objective To colone and analysis of the coding-sequence of p53 gene in U87 glioma cells. Methods We were designed the primers according to the mRNA sequence of p53 from the gene database of NCBI C, enBank. Then we cloned the full-length cDNA sequences of p53 gene from U87 human glioma cells using RT-PCR assay, and compared with the gene database of NCBI C, enBank by using DNAMAN software. And analyze the feature of p53 sequences in U87 glioma cells. Results We cloned the full-length cDNA sequence of p53 gene,and found that it is expression of the wild-type p53 with the Arg genetype at 72 coden in U87 cells. Conclusion We were cloned the 1353 gene and found that it is expression of wild-type p53 gene with the Arg genetype at 72 coden in U87 cells. Our results will be of special help for study the characteristic of p53 gene and the relationship with glioma.