Shore E M, Guild G M
Department of Biology, University of Pennsylvania, Philadelphia 19104-6017.
Genes Dev. 1987 Oct;1(8):829-39. doi: 10.1101/gad.1.8.829.
cis-acting sequence regions involved in the regulation of Sgs-5 gene expression were mapped by testing DNA segments containing the Sgs-5 RNA coding region and various amounts of adjacent sequences for the ability to express Sgs-5 RNA. Following injection of the DNA segments into Drosophila embryos, expression of the gene was assayed in the salivary glands of the injected animals after they developed to third instar larvae, these somatically transformed individuals serving as an in vivo transient expression system. The information necessary for the expression of Sgs-5 is contained within 109 bp upstream and 69 bp downstream of the transcribed region. Somatic transformation experiments also show that some feature within the limits of a 1012-bp DNA segment containing the Sgs-5 RNA coding region derived from the Sgs-5 RNA null stock CA-2 must be responsible for the lack of transcription from this allele. The only DNA sequence differences between active and null alleles, within the 1012 bp, are seven single-base-pair substitutions between -84 bp and +175 bp relative to the RNA start site. One or a combination of these sites are likely contributors to the transcriptional inactivity of the Sgs-5CA2 allele.
通过检测包含Sgs-5 RNA编码区及不同长度相邻序列的DNA片段表达Sgs-5 RNA的能力,对参与Sgs-5基因表达调控的顺式作用序列区域进行了定位。将这些DNA片段注射到果蝇胚胎后,在注射的动物发育至三龄幼虫后,检测其唾液腺中该基因的表达,这些体细胞转化个体作为体内瞬时表达系统。Sgs-5表达所需的信息包含在转录区上游109 bp和下游69 bp范围内。体细胞转化实验还表明,来自Sgs-5 RNA无效品系CA-2的包含Sgs-5 RNA编码区的1012-bp DNA片段范围内的某些特征,必定是该等位基因缺乏转录的原因。在1012 bp范围内,活性等位基因和无效等位基因之间唯一的DNA序列差异是相对于RNA起始位点在-84 bp至+175 bp之间的七个单碱基对替换。这些位点中的一个或组合可能是导致Sgs-5CA2等位基因转录无活性的原因。