Suzuki M G, Shimada T, Kobayashi M
Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Japan.
Heredity (Edinb). 1998 Sep;81 ( Pt 3):275-83. doi: 10.1046/j.1365-2540.1998.00356.x.
To confirm whether sex-linked genes of the silkworm are dosage compensated or not, we investigated three RAPDs and one RFLP marker which have been mapped to the Z chromosome as candidate genes to test for dosage compensation. Northern blot analysis demonstrated that one of the RAPDs, T15.180a, is transcribed into several mRNAs in the embryo, midgut, Malpighian tubule, testis and ovary. Southern blot analysis revealed that T15.180a is present at one copy in the genome. Sequence data showed that T15.180a contained two open-reading frames (ORFs) separated by an intron of 605 bp. These ORFs encoded a possible polypeptide of 445 amino acids containing the proline cluster. These results suggested that T15.180a is part of a functional gene on the Z chromosome of the silkworm. We have called this gene T15, and compared its transcriptional level between males and females. Northern blot analysis showed that the level of T15 mRNA in males was twice that of females. For a more precise analysis, the levels of transcription from both alleles of T15 (T15P, T15C) were measured by the single-nucleotide primer extension (SNuPE) assay. The results indicated that the amount of transcription from each T15 allele in both sexes (male, T15P/T15C; female, T15P/W) was equivalent. All the results suggest that sex-linked genes in the silkworm are not dosage compensated.
为了确定家蚕的性连锁基因是否存在剂量补偿效应,我们研究了三个已定位到Z染色体上作为剂量补偿效应检测候选基因的随机扩增多态性DNA(RAPD)标记和一个限制性片段长度多态性(RFLP)标记。Northern印迹分析表明,其中一个RAPD标记T15.180a在胚胎、中肠、马氏管、睾丸和卵巢中可转录为几种mRNA。Southern印迹分析显示,T15.180a在基因组中以单拷贝形式存在。序列数据表明,T15.180a包含两个开放阅读框(ORF),由一个605 bp的内含子隔开。这些ORF编码一个可能含有脯氨酸簇的445个氨基酸的多肽。这些结果表明,T15.180a是家蚕Z染色体上一个功能基因的一部分。我们将这个基因命名为T15,并比较了其在雄性和雌性之间的转录水平。Northern印迹分析表明,雄性中T15 mRNA的水平是雌性的两倍。为了进行更精确的分析,通过单核苷酸引物延伸(SNuPE)测定法测量了T15两个等位基因(T15P、T15C)的转录水平。结果表明,两性中每个T15等位基因的转录量(雄性,T15P/T15C;雌性,T15P/W)是相等的。所有结果表明,家蚕中的性连锁基因不存在剂量补偿效应。