Monstein H J, Westin G, Philipson L, Pettersson U
EMBO J. 1982;1(1):133-7. doi: 10.1002/j.1460-2075.1982.tb01136.x.
Clones containing sequences complementary to the small nuclear RNA U1 were isolated from the human DNA library of Lawn et al. (1978). Three clones were studied by hybridization and restriction enzyme cleavage. The results showed that the inserts in all three clones were different and that each clone contains one single copy of a sequence which hybridizes to U1 RNA. The results revealed moreover that only one of the three clones contains all the cleavage sites which can be predicted from the known sequence of human U1 RNA, suggesting that the three clones comprise one candidate U1 gene and two pseudogenes. A fragment from the recombinant with the candidate U1 gene was subcloned in the pPR322 plasmid and part of its sequence was determined. The results showed that the subclone contains a sequence which matches that of the human U1 RNA perfectly. The sequence "TATAT" which often is found adjacent to RNA polymerase II start sites, was identified 33-37 base pairs upstream from the beginning of the U1 sequence. Two ten base pairs long, nearly perfect, direct repeats were also identified in the vicinity of the U1 sequence and an imperfect inverted repeat follows immediately after the U1 gene.
从Lawn等人(1978年)构建的人类DNA文库中分离出了含有与小核RNA U1互补序列的克隆。通过杂交和限制性内切酶切割对三个克隆进行了研究。结果表明,所有三个克隆中的插入片段都不同,并且每个克隆都包含一个与U1 RNA杂交的单拷贝序列。此外,结果显示三个克隆中只有一个包含从已知人类U1 RNA序列预测出的所有切割位点,这表明这三个克隆包含一个候选U1基因和两个假基因。将带有候选U1基因的重组体中的一个片段亚克隆到pPR322质粒中,并测定了其部分序列。结果表明,该亚克隆包含一个与人类U1 RNA序列完全匹配的序列。在U1序列起始点上游33 - 37个碱基对处鉴定出了通常位于RNA聚合酶II起始位点附近的“TATAT”序列。在U1序列附近还鉴定出了两个长度为十个碱基对、近乎完美的正向重复序列,并且在U1基因之后紧接着是一个不完美的反向重复序列。