McNabb S, Greig S, Davis T
Department of Genetics, University of Cambridge, England, United Kingdom.
Genetics. 1996 Jun;143(2):897-911. doi: 10.1093/genetics/143.2.897.
This report describes the structure and expression of the outspread (osp) gene of Drosophila melanogaster. Previous work showed that chromosomal breakpoints associated with mutations of the osp locus map to both sides of the alcohol dehydrogenase gene (Adh), suggesting that Adh and the adjacent gene Adhr are nested in osp. We extended a chromosomal walk and mapped additional osp mutations to define the maximum molecular limit of osp as 119 kb. We identified a 6-kb transcript that hybridizes to osp region DNA and is altered or absent in osp mutants. Accumulation of this RNA peaks during embryonic and pupal periods. The osp cDNAs comprise two distinct classes based on alternative splicing patterns. The 5' end of the longest cDNA was extended by PCR amplification. When hybridized to the osp walk, the 5' extension verifies that Adh and Adhr are nested in osp and shows that osp has a transcription unit of > or = 74 kb. In situ hybridization shows that osp is expressed both maternally and zygotically. In the ovary, osp is transcribed in nurse cells and localized in the oocyte. In embryos, expression is most abundant in the developing visceral and somatic musculature.
本报告描述了黑腹果蝇扩展(osp)基因的结构和表达。先前的研究表明,与osp基因座突变相关的染色体断点位于乙醇脱氢酶基因(Adh)两侧,这表明Adh和相邻基因Adhr嵌套在osp中。我们扩展了染色体步移,并对其他osp突变进行定位,以确定osp的最大分子限度为119 kb。我们鉴定出一个6 kb的转录本,它与osp区域DNA杂交,在osp突变体中发生改变或缺失。这种RNA在胚胎期和蛹期积累达到峰值。基于可变剪接模式,osp cDNA可分为两个不同的类别。通过PCR扩增延伸了最长cDNA的5'端。当与osp步移杂交时,5'端延伸证实Adh和Adhr嵌套在osp中,并表明osp有一个≥74 kb的转录单元。原位杂交显示osp在母体和合子期均有表达。在卵巢中,osp在滋养细胞中转录并定位在卵母细胞中。在胚胎中,表达在发育中的内脏和体壁肌肉组织中最为丰富。