Sassone-Corsi P, Corden J, Kédinger C, Chambon P
Nucleic Acids Res. 1981 Aug 25;9(16):3941-58. doi: 10.1093/nar/9.16.3941.
We have cloned into plasmid pBR322 a DNA fragment extending from position -32 to position -12 of the adenovirus type 2 major late promoter region (position +1 referring to the cap site). In vitro transcription experiments show that this 21 base pair sequence, which contains the Goldberg-Hogness or "TATA" box, is both necessary and sufficient for specific initiation of transcription by RNA polymerase B (or II). Furthermore, we show that similar sequences, randomly occurring in the bacterial plasmid pBR322, are also recognized by the RNA polymerase B transcription machinery and able to promote specific in vitro transcription. Finally, we discuss the possible importance of the nucleotide sequence of the start region in the actual efficiency of initiation of in vitro transcription.
我们已将一段从腺病毒2型主要晚期启动子区域的-32位延伸至-12位的DNA片段(+1位指帽位点)克隆到质粒pBR322中。体外转录实验表明,这段包含戈德堡-霍格尼斯盒或“TATA”盒的21个碱基对序列,对于RNA聚合酶B(或II)特异性起始转录而言,既是必需的也是充分的。此外,我们还表明,随机存在于细菌质粒pBR322中的类似序列,也能被RNA聚合酶B转录机制识别,并能促进特异性体外转录。最后,我们讨论了起始区域核苷酸序列在体外转录起始实际效率中的可能重要性。