Shen C K, Maniatis T
J Mol Appl Genet. 1982;1(4):343-60.
We have studied the location, structure, and in vitro transcription of repetitive DNA sequences within the human alpha-like globin gene cluster. At least eight different Alu family repeats were identified, each of which is transcribed in vitro to produce discrete RNA transcripts. The nucleotide sequence of one Alu repeat sequence, located on the 3' side of the alpha l globin gene (3'-alpha l), was determined and compared to published Alu repeat sequences. In vitro transcription of this repeat sequence generates RNA fragments of approximately 410, 260, 160, and 86 nucleotides. To determine whether these transcripts associate with specific proteins in vitro, we carried out immunoprecipitation experiments using an antiserum from systemic lupus erythematosus (SLE) patients. We find that the antiserum anti-La, which was shown to precipitate ribonucleoproteins (RNPs) containing the adenovirus VAI RNA from virus infected cells, preferentially precipitates the smallest two in vitro transcripts of the 3'-alpha l Alu repeat. These results suggest that the RNAs interact with specific factors in the in vitro transcription reaction mix to form RNP.
我们研究了人类α-珠蛋白基因簇中重复DNA序列的定位、结构及体外转录情况。至少鉴定出了八种不同的Alu家族重复序列,每个重复序列在体外均可转录产生离散的RNA转录本。确定了位于α1珠蛋白基因(3'-α1)3'侧的一个Alu重复序列的核苷酸序列,并与已发表的Alu重复序列进行了比较。该重复序列的体外转录产生了约410、260、160和86个核苷酸的RNA片段。为了确定这些转录本在体外是否与特定蛋白质结合,我们使用系统性红斑狼疮(SLE)患者的抗血清进行了免疫沉淀实验。我们发现,抗La抗血清(已证明可从病毒感染细胞中沉淀出含有腺病毒VAI RNA的核糖核蛋白(RNP))优先沉淀3'-α1 Alu重复序列的最小的两个体外转录本。这些结果表明,RNA在体外转录反应混合物中与特定因子相互作用形成RNP。