DIFFERENTIAL SCREENING OF A HUMAN PANCREATIC ADENOCARCINOMA LAMBDA-GT11 EXPRESSION LIBRARY HAS IDENTIFIED INCREASED TRANSCRIPTION OF ELONGATION-FACTOR EF-1-ALPHA IN TUMOR-CELLS
DIFFERENTIAL SCREENING OF A HUMAN PANCREATIC ADENOCARCINOMA LAMBDA-GT11 EXPRESSION LIBRARY HAS IDENTIFIED INCREASED TRANSCRIPTION OF ELONGATION-FACTOR EF-1-ALPHA IN TUMOR-CELLS
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DOI：
10.1002/ijc.2910500513
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发表时间：
1992-03-12
影响因子：
6.4
通讯作者：
GRANT, DAW
中科院分区：
文献类型：
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作者：
GRANT, AG;FLOMEN, RM;GRANT, DAW
使用来自正常和癌性胰腺组织的 mRNA 差异筛选人胰腺腺癌 lambda-gt11 表达文库。五个克隆优先与胰腺腺癌 mRNA 杂交。通过 PCR 扩增来自其中 4 个克隆的 cDNA 插入片段，用 alpha-P-32 标记，并用于针对从多种肿瘤和正常组织制备的 mRNA 进行 Northern 印迹分析。 Lambda-GER-4 鉴定出一种胰腺相关 mRNA (> 10 kb)，在核酸或氨基酸水平上与已知序列没有同源性。 Lambda-GER-2 鉴定出一个 1.7 kb mRNA 转录物，相对于正常组织，该转录物在从胰腺、结肠、乳腺、肺和胃肿瘤制备的 mRNA 中过度表达。序列分析和酶切图谱显示该克隆与人延伸因子EF-1-α的活性形式完全同源。肿瘤组织中 EF-1-α-mRNA 的高水平表达为越来越多的证据表明 EF-1-α 是细胞周期的重要调节因子提供了支持。
A human pancreatic adenocarcinoma lambda-gt11 expression library was differentially screened with mRNA derived from normal and cancerous pancreatic tissues. Five clones preferentially hybridized with pancreatic adenocarcinoma mRNA. cDNA inserts from 4 of these clones were amplified by PCR, labelled with alpha-P-32 and used in Northern blot analysis against mRNA prepared from a variety of tumour and normal tissues. Lambda-GER-4 identified a pancreas-associated mRNA (> 10 kb) with no homology with known sequences at either the nucleic or amino-acid level. Lambda-GER-2 identified a 1.7-kb mRNA transcript that was over-expressed in mRNA prepared from pancreas, colon, breast, lung and gastric tumours relative to normal tissues. Sequence analysis and restriction-enzyme mapping showed that this clone was completely homologous with the active form of human elongation factor EF-1-alpha. This high level of EF-1-alpha-mRNA expression in tumour tissues lends support to the increasing evidence that EF-1-alpha is an important regulator of the cell cycle.