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在瞬时分析中珠蛋白基因转录的顺式和反式激活

cis and trans activation of globin gene transcription in transient assays.

作者信息

Treisman R, Green M R, Maniatis T

出版信息

Proc Natl Acad Sci U S A. 1983 Dec;80(24):7428-32. doi: 10.1073/pnas.80.24.7428.

Abstract

We examined the effects of the simian virus 40 enhancer sequence on transcription of cloned human alpha- and beta-globin genes shortly after their introduction into cultured mammalian cells. We find that (i) detectable transcription of the beta-globin gene but not the alpha-globin gene requires linkage to the enhancer; (ii) the enhancer increases the amount of beta-globin RNA at least 100-fold but results in only a 5- to 10-fold increase in the amount of alpha-globin RNA; (iii) plasmid replication does not increase the level of beta-globin RNA, regardless of linkage to the enhancer, but does result in an approximately equal to 50-fold increase in the level of alpha-globin RNA; (iv) the enhancer is not required for and does not increase transcription of either gene in 293 cells, an adenovirus 5-transformed human kidney cell line. We also show that an enhancer sequence is not required for activity of the normally enhancer-dependent simian virus 40 early promoter in 293 cells, indicating that these cells contain a trans-acting factor(s) that circumvents the requirement for the enhancer sequence.

摘要

在将克隆的人α-和β-珠蛋白基因导入培养的哺乳动物细胞后不久,我们检测了猿猴病毒40增强子序列对其转录的影响。我们发现:(i)β-珠蛋白基因可检测到的转录,但α-珠蛋白基因则不然,这需要与增强子相连;(ii)增强子使β-珠蛋白RNA的量增加至少100倍,但α-珠蛋白RNA的量仅增加5至10倍;(iii)质粒复制不会增加β-珠蛋白RNA的水平,无论是否与增强子相连,但确实会使α-珠蛋白RNA的水平增加约50倍;(iv)在293细胞(一种腺病毒5转化的人肾细胞系)中,增强子对于这两个基因的转录既不是必需的,也不会增加其转录。我们还表明,在293细胞中,正常依赖增强子的猿猴病毒40早期启动子的活性不需要增强子序列,这表明这些细胞含有一种反式作用因子,可绕过对增强子序列的需求。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8eb/389964/099cbafcb871/pnas00650-0062-a.jpg

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