EXPRESSION OF A BETA-GLOBIN GENE IS ENHANCED BY REMOTE SV40 DNA-SEQUENCES
EXPRESSION OF A BETA-GLOBIN GENE IS ENHANCED BY REMOTE SV40 DNA-SEQUENCES
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DOI：
10.1016/0092-8674(81)90413-x
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发表时间：
1981-01-01
期刊：
影响因子：
64.5
通讯作者：
SCHAFFNER, W
中科院分区：
文献类型：
--
作者：
BANERJI, J;RUSCONI, S;SCHAFFNER, W
研究了克隆的兔Hb β 1基因导入人宫颈癌HeLa细胞后的瞬时表达。在转染后2.5天，使用磷酸钙技术，从整个细胞群中提取RNA，并通过S1核酸酶杂交测定进行分析。当β-使用珠蛋白基因-质粒重组体。然而，200倍以上的β-当β-珠蛋白基因转录物被发现时，珠蛋白基因重组体也含有SV 40 DNA和90%的这些转录物（约1000个/细胞）具有与真实兔珠蛋白mRNA相同的5“末端。大量生产β-通过免疫荧光染色，在转染细胞的一部分中容易地检测到珠蛋白。珠蛋白基因表达的增强依赖于SV 40序列的顺式作用，但独立于病毒的DNA复制起点。增强活性与位于病毒晚期基因区开始处的72个碱基对（bp）重复序列元件相关。含有转录增强子元件的病毒DNA片段可以在任一方向上在许多位置起作用，包括兔β-CD的转录起始位点上游1400 bp或下游3300 bp。珠蛋白基因定义了一类具有迄今为止未描述的作用模式的DNA元件。通过特异性增强子元件激活基因似乎是一种广泛的机制，可用于调节基因表达。
The transient expression of a cloned rabbit Hb .beta.1 gene was studied after its introduction into human cervical carcinoma HeLa cells. At 2.5 days after transfection, using the calcium phosphate technique, RNA was extracted from the entire cell population and analyzed by the S1 nuclease hybridization assay. Transcripts were barely detectable when .beta.-globin gene-plasmid recombinants were used. However, 200 times more .beta.-globin gene transcripts were found when the .beta.-globin gene recombinants also contained SV40 DNA and 90% of these transcripts (.apprx. 1000/cell) had the same 5'' end as authentic rabbit globin mRNA. Abundant production of .beta.-globin protein was readily detected in a fraction of transfected cells by immunofluorescent staining. Enhancement of globin gene expression was dependent on SV40 sequences acting in cis, but independent of the viral origin of DNA replication. The enhancing activity was associated with the 72 base pair (bp) repeated sequence element located at the beginning of the viral late gene region. Viral DNA fragments containing the transcriptional enhancer element could act in either orientation at many positions, including 1400 bp upstream or 3300 bp downstream from the transcription initiation site of the rabbit .beta.-globin gene. A class of DNA elements with a mode of action that was not heretofore described is defined. The activation of genes by specific enhancer elements seems to be a widespread mechanism that may be used for the regulation of gene expression.