家蚕Ring基因的克隆及序列特征与组织表达分析

发布时间：2018-10-31 20:44

【摘要】：Polycomb group(PcG)蛋白是多细胞有机体中的一类表观遗传抑制因子,Ring是Pc G蛋白家族的主要成员,在Pc G蛋白的转录抑制调控中行使重要功能,对昆虫和哺乳动物的发育至关重要。利用同源检索方法获得家蚕Ring基因(BmRing)的核苷酸序列,该基因位于家蚕第17号染色体的nscaf2865位点。BmRing的核苷酸序列由4个外显子和3个内含子构成,基因的c DNA3'端有2个多聚腺苷酸化信号(aataaa)和典型的poly(A)结构;BmRing编码由377个氨基酸组成的蛋白质,分子质量为41.8 k D,等电点为6.72,氨基酸序列具有3个同源异型结构域(homeobox domain,HD),46~85 aa为高度保守的典型的环指结构域(Ring finger),331~347 aa为跨膜区,N端位于膜外。通过STRING在线预测分析得到10个与BmRing有相互作用关系的蛋白质。系统进化分析显示BmRing与果蝇的Dm SCE、意大利蜜蜂的AmRing等的亲缘关系较近。以家蚕5龄第3天幼虫的卵巢c DNA为模板,克隆了BmRing基因,并利用半定量RT-PCR检测BmRing基因在5龄第3天幼虫的精巢、卵巢、头部和血细胞中有明显表达,而在其他组织中几乎不表达。获得的上述基础信息,有益于进一步研究BmRing的生物学功能。
[Abstract]:Polycomb group (PcG) protein is an epigenetic inhibitor in multicellular organisms. Ring is a major member of Pc G protein family. It plays an important role in the regulation of transcription inhibition of Pc G protein and plays an important role in the development of insects and mammals. The nucleotide sequence of Ring gene (BmRing) of Bombyx mori was obtained by homology search method. The nucleotide sequence of BmRing gene was composed of 4 exons and 3 introns, which was located at the nscaf2865 site of chromosome 17 of Bombyx mori. There are two polyadenylate signal (aataaa) and typical poly (A) structure at the c DNA3' end of the gene. BmRing encodes a 377 amino acid protein with a molecular weight of 41.8 KD and a isoelectric point of 6.72. The amino acid sequence has three homologous domains (homeobox domain,HD). 46 ~ 85 aa is a typical conserved ring finger domain, (Ring finger), 331n 347 aa is a transmembrane region, and the N terminal is located outside the membrane. Ten proteins with interaction with BmRing were obtained by STRING online prediction analysis. Phylogenetic analysis showed that BmRing was closely related to Dm SCE, of Drosophila melanogaster Dm SCE, AmRing et al. The BmRing gene was cloned from the ovary c DNA of the 3rd day larva of the 5th instar of Bombyx mori. The expression of BmRing gene was detected by semi-quantitative RT-PCR in the spermary, ovary, head and blood cells of the 3rd day larva of the 5th instar. In other tissues there is little expression. The above basic information is helpful to further study the biological function of BmRing.
【作者单位】：江苏科技大学生物技术学院;家蚕基因组生物学国家重点实验室西南大学;
【基金】：国家自然科学基金项目(No.31302035) 江苏省高校自然科学基金项目(No.13KJD230001) 江苏省自然科学基金项目(No.BK2012273) 江苏科技大学引进人才科研启动基金项目(No.35181005,35181003)
【分类号】：S881.2;Q78

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