Fasano Laurent, Coré Nathalie, Kerridge Stephen
LGBC, CNRS, Centre Universitaire de Marseille-Luminy, Case 907, F-13288, Marseille Cedex 9, France.
Rouxs Arch Dev Biol. 1988 Jan;197(8):507-512. doi: 10.1007/BF00385685.
Random insertions of a promotor fused to a reporter gene, such as Lac-Z, reveal regulatory sequences that confer temporal and spatial patterns of gene expression in eukaryotes. These patterns may reflect the activity of a neighbouring gene and thus lead to the isolation of new genes essential for normal development. Here, we demonstrate that this hypothesis is true for an insertion into the well characterized segmentation gene, hairy, in Drosophila. The insertion is homozygous lethal and fails to complement other hairy alleles, giving the phenotype described for hairy mutations. The insertion is located at 66D on the polytene chromosome map, is within 300-600 bp 5' to the first hairy exon, and is orientated in the same sense (5'-3') as the hairy transcription unit. Expression of β-galactosidase (β-gal), deriving from the insertion, follows closely the spatio-temporal patterns of expression of hairy gene product during embryogenesis. In addition, other sites of β-galactosidase expression are shown in the third larval instar stage and in the adult ovary. The results show that some insertions, giving restricted patterns of reporter gene expression, will reflect the temporo-spatial activity of a nearby gene.
将与报告基因(如Lac-Z)融合的启动子随机插入,可揭示在真核生物中赋予基因表达时空模式的调控序列。这些模式可能反映相邻基因的活性,从而导致分离出正常发育所必需的新基因。在此,我们证明这一假设对于果蝇中插入到特征明确的分节基因hairy中的情况是正确的。该插入是纯合致死的,并且不能与其他hairy等位基因互补,呈现出hairy突变所描述的表型。该插入位于多线染色体图谱的66D处,在第一个hairy外显子的5'端300 - 600 bp范围内,并且与hairy转录单元的方向相同(5'-3')。源自该插入的β-半乳糖苷酶(β-gal)的表达在胚胎发育过程中紧密跟随hairy基因产物的时空表达模式。此外,在第三龄幼虫阶段和成虫卵巢中也显示出β-半乳糖苷酶表达的其他位点。结果表明,一些导致报告基因表达模式受限的插入将反映附近基因的时空活性。