Identification of two dazl splicing variants in Japanese flounder (Paralichthys olivaceus)
首发时间:2016-05-23
Abstract:DAZL is an RNA-binding protein of DAZ family, playing key roles in reproduction of organisms. Here in this study, we clone and identify the dazl gene in a kind of flatfish, Japanese flounder (Paralichthys olivaceus). Two different splicing variants of dazl mRNAs are found, and they differ from each other by a 51-bp insertion/deletion. We characterize the two mRNA variants by qPCR and in situ hybridization and prove their germ cell specific expression patterns. Results of western blot and immunohistochemistry show that the two isoforms of their translational production also express specifically in germ cells of both sexes. The similar expression patterns of the two variants may imply their complementary functions in germ cell development. But there are also differences in their functions. By RNA-binding protein immunoprecipitation, we find Podazl-I and Podazl-II may be the target of both PODAZL isoforms. More sequencing results will offer more detailed and accurate information on their functions. Studies on the two dazl variants in Japanese flounder are rare in teleosts. Our results may facilitate further studies on reproduction related genes and germ cell development.
keywords: Genetics dazl RNA binding protein alternative splice germ cell Japanese flounder
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牙鲆DAZL两种可变剪接形式的鉴定及功能研究
摘要:DAZL是DAZ家族的成员之一,同时也是一种重要的RNA结合蛋白。现有研究发现,从无脊椎动物到脊椎动物,dazl特异地表达于雌性和雄性生殖细胞,可能在生殖细胞发育过程发挥重要作用。本研究克隆和鉴定了牙鲆(Paralichthys olivaceus) dazl基因,首次在硬骨鱼中发现了其mRNA的可变剪接形式,将这两种牙鲆dazl mRNA称为Podazl-I和Podazl-II,探究了它们在mRNA水平和蛋白水平的表达规律,并找到了牙鲆DAZL的靶mRNA,预测其功能。牙鲆dazl两种mRNA和蛋白形式的表达规律比较及靶mRNA比较结果说明,两者在功能上可能不存在明显差异,或许牙鲆dazl较短的可变剪接形式的出现可以更高效的进行转录和翻译,与完整的剪接形式一起,共同执行相似的生物学功能。这些结果为牙鲆及其它硬骨鱼的生殖和发育研究提供重要的参考依据,为种质资源的利用和保存、育种工作的开展奠定基础。
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No.4689150523082146****
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