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首页> 外文期刊>African Journal of Biotechnology >Construction and characterization of a cDNA library from human brain glioma cell line U251 with overexpressed exogenous p53 gene
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Construction and characterization of a cDNA library from human brain glioma cell line U251 with overexpressed exogenous p53 gene

机译：人脑胶质瘤细胞系U251中过表达外源p53基因cDNA文库的构建与鉴定

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The tumor-suppressor gene p53 and its downstream genes consist of a complicated gene network, and the challenge to understand the network is to identify p53 downstream genes. In order to isolate and identify new p53 regulated genes, we constructed and characterized a normalized cDNA library from human brain glioma cell line U251 while exogenous p53 gene is overexpressed. The constructed cDNA library contained 1.3×106?directional recombinants, and its insert size ranged from 0.5 to 2.0 kb. Screening the cDNA library, we obtained two novel p53 downstream genes, PAP1 and PAP2. Polymerase chain reaction (PCR) analyses of the library for specific genes revealed the presence of cDNAs for p53 downstream genes such as p21, gadd45, and PCNA. These results demonstrate the sequence complexity and relatively low redundancy of our cDNA library. It is a valuable and unique resource for studying p53 gene expression, regulatory mechanisms and screening p53 downstream genes.
机译：肿瘤抑制基因p53及其下游基因由一个复杂的基因网络组成，要理解该网络的挑战是鉴定p53下游基因。为了分离和鉴定新的p53调控基因，我们构建并表征了人脑神经胶质瘤细胞系U251中归一化的cDNA文库，而外源p53基因却过表达。所构建的cDNA文库包含1.3×106′方向的重组体，其插入大小为0.5至2.0kb。筛选cDNA文库，我们获得了两个新的p53下游基因PAP1和PAP2。文库中针对特定基因的聚合酶链反应（PCR）分析显示，存在p53下游基因（例如p21，gadd45和PCNA）的cDNA。这些结果证明了我们cDNA文库的序列复杂性和相对较低的冗余度。它是研究p53基因表达，调控机制和筛选p53下游基因的宝贵且独特的资源。

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