Miller W G, Simons R W
Department of Microbiology, University of California, Los Angeles 90024.
Mol Microbiol. 1990 Jun;4(6):881-93. doi: 10.1111/j.1365-2958.1990.tb00661.x.
We present evidence that DNA from diverse prokaryotic and eukaryotic sources gives rise to low-level fusion expression in Escherichia coli promoter-probe vectors. This expression may be as high as approximately 10% of the E. coli lacUV5 promoter. Although expression does not correlate with the presence of obvious E. coli promoter-like sequences, it is blocked by transcriptional terminators. Furthermore, transcription across the fusion junction is detected at levels that correlate with fusion expression. We suggest that this 'low-level transcription' (LLT) results from infrequent initiation by RNA polymerase at random sites and/or weak promoters. We propose that LLT has biological significance. In some instances, it may provide an advantageous basal level of gene expression, and we suggest that this may be true for the E. coli lacY gene. In other instances, LLT may be detrimental, in which case it may be blocked by mechanisms such as RNA secondary structure or transcriptional polarity. We present evidence to show that activation of the IS10 transposase gene by LLT is blocked at the translational level.
我们提供的证据表明,来自不同原核生物和真核生物来源的DNA在大肠杆菌启动子探针载体中会引发低水平的融合表达。这种表达可能高达大肠杆菌lacUV5启动子的约10%。尽管表达与明显的大肠杆菌启动子样序列的存在无关,但它会被转录终止子阻断。此外,在与融合表达相关的水平上检测到跨越融合接头的转录。我们认为这种“低水平转录”(LLT)是由RNA聚合酶在随机位点和/或弱启动子处的不频繁起始导致的。我们提出LLT具有生物学意义。在某些情况下,它可能提供有利的基因表达基础水平,我们认为大肠杆菌lacY基因可能就是这种情况。在其他情况下,LLT可能是有害的,在这种情况下,它可能会被诸如RNA二级结构或转录极性等机制阻断。我们提供证据表明,LLT对IS10转座酶基因的激活在翻译水平上被阻断。