Honda B M, Devlin R H, Nelson D W, Khosla M
Nucleic Acids Res. 1986 Jan 24;14(2):869-81. doi: 10.1093/nar/14.2.869.
Using the nematode Caenorhabditis elegans as a model organism, we have prepared cell-free extracts which accurately transcribe cloned homologous 5S RNA genes in vitro. These extracts also transcribe cloned tRNA genes, and actively process the resulting products. Unlike tRNA genes, transcription of 5S DNA shows some species specificity: C. elegans extracts do not transcribe Xenopus 5S RNA genes, nor does a Xenopus extract efficiently transcribe the heterologous nematode 5S DNA. However, addition of limiting amounts of C. elegans fractions permits Xenopus extracts to transcribe nematode 5S RNA genes. This apparent biochemical "complementation" may provide an assay to purify 5S RNA gene-specific factors from C. elegans.
我们以线虫秀丽隐杆线虫作为模式生物，制备了无细胞提取物，该提取物能够在体外准确转录克隆的同源5S RNA基因。这些提取物还能转录克隆的tRNA基因，并对产生的产物进行活性加工。与tRNA基因不同，5S DNA的转录表现出一定的物种特异性：秀丽隐杆线虫提取物不能转录非洲爪蟾的5S RNA基因，非洲爪蟾提取物也不能有效地转录异源线虫的5S DNA。然而，添加有限量的秀丽隐杆线虫组分可使非洲爪蟾提取物转录线虫的5S RNA基因。这种明显的生化“互补”可能为从秀丽隐杆线虫中纯化5S RNA基因特异性因子提供一种检测方法。
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