Stebbings L, Grimes B R, Bownes M
The University of Edinburgh, Institute of Cell and Molecular Biology, Darwin Building, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.
Dev Genes Evol. 1998 Nov;208(9):523-30. doi: 10.1007/s004270050211.
Differential screening was used to isolate a genomic clone, lambda89Ba located at 89B that hybridised preferentially with female cDNA. On further investigation, a 3.1-kb subfragment, 89Ba(3.1), was shown to contain a gene with male germline-specific expression (Mst89B) flanked by two genes (Mat89Ba and Mat89Bb) expressed predominantly in the ovaries and embryo of Drosophila melanogaster. Mat89Bb is separated from Mst89B by at most 100 bp; Mst89B and Mat89Ba are convergently transcribed and their 3' untranslated regions (UTRs) overlap by a minimum of 85 bp. Database searches with either the 89Ba(3.1) genomic DNA sequence or conceptual translations of Mst89B or Mat89Bb cDNAs failed to reveal any significant similarities with database entries. Using in situ hybridisation to ovaries, Mat89Ba and Mat89Bb were shown to be expressed in nurse cells at stages nine and ten of oogenesis and exported to the oocyte. In addition, Mat89Bb transcripts were detected in the follicle cells surrounding the oocyte. Mst89B transcripts were present throughout spermatogenesis in germline-derived cells, consistent with northern analysis which showed that they were absent in the offspring of tudor flies that lack a germline. The absence of Mst89B transcripts at the tip of the testis suggested that the somatic cells in this region do not express Mst89B. Two 12-bp sequences were identified in the 5' UTR of Mst89B with a strong similarity to translational control elements (TCEs) originally identified in the CGP gene family. This suggests that TCEs may be present in a wider class of testis transcripts.
采用差异筛选法分离出一个位于89B的基因组克隆lambda89Ba,它优先与雌性cDNA杂交。进一步研究表明,一个3.1 kb的亚片段89Ba(3.1)包含一个具有雄性生殖系特异性表达的基因(Mst89B),其两侧是两个主要在黑腹果蝇卵巢和胚胎中表达的基因(Mat89Ba和Mat89Bb)。Mat89Bb与Mst89B的间隔最多为100 bp;Mst89B和Mat89Ba反向转录,它们的3'非翻译区(UTR)至少重叠85 bp。用89Ba(3.1)基因组DNA序列或Mst89B或Mat89Bb cDNA的概念性翻译进行数据库搜索,均未发现与数据库条目有任何显著相似性。通过对卵巢进行原位杂交,发现Mat89Ba和Mat89Bb在卵子发生的第九和第十阶段在滋养细胞中表达,并输出到卵母细胞中。此外,在卵母细胞周围的卵泡细胞中检测到Mat89Bb转录本。Mst89B转录本在生殖系来源的细胞整个精子发生过程中都存在,这与Northern分析结果一致,Northern分析表明它们在缺乏生殖系的tudor果蝇的后代中不存在。睾丸顶端缺乏Mst89B转录本,表明该区域的体细胞不表达Mst89B。在Mst89B的5'UTR中鉴定出两个12 bp的序列,与最初在CGP基因家族中鉴定的翻译控制元件(TCE)有很强的相似性。这表明TCE可能存在于更广泛的睾丸转录本类别中。