Olsson P, Bera T K, Essand M, Kumar V, Duray P, Vincent J, Lee B, Pastan I
Laboratory of Molecular Biology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-4255, USA.
Prostate. 2001 Sep 15;48(4):231-41. doi: 10.1002/pros.1102.
The database of human expressed sequence tags (dbEST) is a potential source for the identification of tissue specific genes. The database contains sequences that originate from cDNA libraries from different tissues cell types and tumors.
Computer based analysis identified a cluster of sequence homologous ESTs, containing ESTs derived only from human prostate cDNA libraries. The tissue specificity was examined by multiple tissue RNA dot blots and RT-PCR. The new RNA transcript was characterized using northern blot analysis, RACE-PCR, and a ribonuclease protection assay.
We have identified a gene differentially expressed in prostate using EST database analysis and experimental studies. We name the gene GDEP for gene differentially expressed in prostate. The major GDEP transcript is about 520 bp long. GDEP RNA was detected in nine prostate tissue samples, four normal and five cancer. Expression in prostate epithelial cells was established by in situ hybridization. Weak expression was detected in the prostate cancer cell line LNCaP. In vitro transcription/translation indicate that the RNA encodes a small 34 amino acid protein. The major transcript consists of two exons with one large intron (> 15 kb). The GDEP gene was mapped to chromosome 4q21.1 by radiation hybrid mapping.
Our data proves that tissue specific genes can be identified by EST database mining. The prostate specificity of GDEP expression indicates that GDEP may be useful in the diagnosis or treatment of prostate cancer. Published 2001 Wiley-Liss, Inc.
人类表达序列标签数据库(dbEST)是鉴定组织特异性基因的潜在来源。该数据库包含源自不同组织、细胞类型和肿瘤的cDNA文库的序列。
基于计算机的分析鉴定出一组序列同源的EST,其中仅包含源自人前列腺cDNA文库的EST。通过多组织RNA斑点印迹和RT-PCR检测组织特异性。使用Northern印迹分析、RACE-PCR和核糖核酸酶保护试验对新的RNA转录本进行表征。
我们通过EST数据库分析和实验研究鉴定出一个在前列腺中差异表达的基因。我们将该基因命名为GDEP,即前列腺中差异表达的基因。GDEP的主要转录本约520bp长。在9个前列腺组织样本中检测到GDEP RNA,其中4个为正常样本,5个为癌组织样本。通过原位杂交确定其在前列腺上皮细胞中的表达。在前列腺癌细胞系LNCaP中检测到弱表达。体外转录/翻译表明该RNA编码一种34个氨基酸的小蛋白。主要转录本由两个外显子和一个大内含子(>15kb)组成。通过辐射杂种图谱分析将GDEP基因定位于4q21.1染色体。
我们的数据证明可以通过挖掘EST数据库来鉴定组织特异性基因。GDEP表达的前列腺特异性表明GDEP可能在前列腺癌的诊断或治疗中有用。2001年由Wiley-Liss公司出版。