Construction and characterization of a cDNA library from human brain glioma cell line U251 with overexpressed exogenous p53 gene

  • Kun-xian Shu
  • Yan-zhe Li
  • Hong-bo Shao
  • Ming-hua Zhang
  • Li-xiang Wu

Abstract

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.

Keywords: p53 Gene, p53 downstream gene, cDNA library, normalization

African Journal of Biotechnology Vol. 9(33), pp. 5262-5268, 16 August, 2010

Author Biographies

Kun-xian Shu
College of Bio-information, Chongqing University of Posts and Telecommunications, Chongqing 400065, China
Yan-zhe Li
College of Bio-information, Chongqing University of Posts and Telecommunications, Chongqing 400065, China
Hong-bo Shao
Institute of Soil and Water Conservation, Chinese Academy of Sciences and Ministry of Water Resources , Yangling 712100, China; Yantai Institute of Costal Zone Research, Chinese Academy of Sciences, Yantai 264003, China; Institute for Life Sciences, Qingdao University of Science and Technology, Qingdao 266042, China
Ming-hua Zhang
Xiangya School of Medicine, Central South University, Changsha 410008, China
Li-xiang Wu
Xiangya School of Medicine, Central South University, Changsha 410008, China
Published
2013-08-08
Section
Articles

Journal Identifiers


eISSN: 1684-5315