Funaguma Shunsuke, Hashimoto Shin-ichi, Suzuki Yutaka, Omuro Naoko, Sugano Sumio, Mita Kazuei, Katsuma Susumu, Shimada Toru
Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Insect Biochem Mol Biol. 2007 Feb;37(2):147-54. doi: 10.1016/j.ibmb.2006.11.001. Epub 2006 Nov 15.
To identify genes involved in the differentiation of Bombyx cystoblast, we constructed two 3' long serial analysis of gene expression (Long SAGE) libraries from stage 1-3 or stage 2-3 egg chambers and compared their gene expression profiles. In both libraries, the most frequent tags were derived from the same novel transcript. The transcript does not have any open reading frame capable of encoding a protein with over 100 amino acids in length. RNA blot analysis revealed that this transcript is specifically and abundantly expressed in the Bombyx ovary, mainly the germ line cells in the ovarioles. These results suggest that Bombyx oogenesis may be regulated by a previously unidentified non-coding RNA. Comparison of the gene expression profiles between the stage 1-3 and stage 2-3 egg chamber libraries revealed that 272 tags were significantly more abundant in stage 1-3 egg chambers (p<0.05 and at least two-fold change) than in library 2. Among the differentially expressed transcripts were the sequences that correspond to ATP synthase subunit d (3.1-fold enriched) and ATP synthase coupling factor 6 (9.1-fold enriched), suggesting that they are involved in regulation of cell cycle of cystocytes.
为了鉴定参与家蚕成囊细胞分化的基因,我们构建了两个3'端基因表达系列分析(Long SAGE)文库,分别来自1-3期或2-3期的卵室,并比较了它们的基因表达谱。在这两个文库中,最常见的标签都来自同一个新转录本。该转录本没有任何能够编码长度超过100个氨基酸的蛋白质的开放阅读框。RNA印迹分析表明,这个转录本在家蚕卵巢中特异性且大量表达,主要在卵巢管中的生殖系细胞中表达。这些结果表明,家蚕卵子发生可能受一种先前未鉴定的非编码RNA调控。比较1-3期和2-3期卵室文库之间的基因表达谱发现,272个标签在1-3期卵室中比在文库2中显著更丰富(p<0.05且至少有两倍的变化)。差异表达的转录本中包括与ATP合酶亚基d(富集3.1倍)和ATP合酶偶联因子6(富集9.1倍)相对应的序列,这表明它们参与了成囊细胞细胞周期的调控。