Phylogenetic distribution and expression of amelogenin genes in vertebrates

脊椎动物牙釉蛋白基因的系统发育分布和表达

• 批准号: 07671993
• 负责人: ISHIYAMA Mikio
• 金额: $ 0.58万
• 依托单位: The Nippon Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671993/
• 关键词: amelogenin; gene structure; DNA; Southern hybridyzation; PCR; phylogeny; エナメル蛋白; エナメル質進化; サザンハイブリダイゼーション; 遺伝子発現; in situハイブリダイゼーション

In order to understand the phylogenetic distribution and expression of amelogenin genes in vertebrates, Southern hybridyzation, in situ hybridyzation and PCR amplification were performed in thirteen vertebrate species selected from seven taxonomic classes. The species used in this study were as follows ; the human, mouse (Mammalia), chicken (Aves), caiman, tortoise, snake (Reptilia), frog (Amphibia), gar-pike, lungfish, eel (Osteichthyes), dogfish (Chondrichthyes) and hagfish, lamprey (Agnatha). The Southern hybridyzation assay revealed that the hybridyzation with the human cDNA non-radioactive probes was obtained in the genomic DNA of all the species used. This demonstrates partial DNA sequence similarities between these species, and may indicates the universal distribution of the amelogenin gene or amelogenin gene-like sequences in vertebrates. Then, the gar-pike was selected and was examined by in situ hybridyzation by using human cDNA non-radioactive probes, because of it's signifi … More cance for the phylogenetical location on the enamel evolutions as revealed by fine-structural and immunocytochemical analyzes. Although the hybridyzation was detected with the human cDNA probes by Southern hybridyzation, the signals of mRNA was not detected in the tooth germs of the gar-pike. Furthermore, in the PCR amplification targeted to the homologous sequences with the exon 6 of human amelogenin gene, the products were obtained in the genomic DNA of the human (400bp), mouse (400bp), chicken (780bp), caiman (400bp), tortoise (750bp) and snake (750bp). However no products were obtained in frogs, fishes and cyclostomes. After performing the sequencing of products in the chicken, caiman, tortoise and snake, the DNA sequences of the caiman only corresponded with the exon 6 of human amelogenin gene by 48.5%. A lower but slight homology were recognized in the products of the chicken, tortoiose and snake. Consequently, the present study demonstrates that the amelogenin gene of amelogenin gene-like sequences may be conserved universally in vertebrates. Less

为了了解淀粉原蛋白基因在脊椎动物中的系统发育分布和表达，对7个分类纲的13种脊椎动物进行了Southern杂交、原位杂交和PCR扩增。本研究使用的树种如下：人类、老鼠（哺乳目）、鸡（鸟类）、凯门鳄、乌龟、蛇（爬行目）、青蛙（两栖目）、梭子鱼、肺鱼、鳗鱼（骨鱼目）、角鲨（软骨鱼目）和盲鳗、七鳃鳗（Agnatha）。南方杂交实验表明，在所有使用的物种基因组DNA中都获得了与人类cDNA非放射性探针的杂交。这表明这些物种之间的部分DNA序列相似，并可能表明淀粉原蛋白基因或淀粉原蛋白基因样序列在脊椎动物中的普遍分布。由于其在牙釉质进化过程中具有重要的系统发育位置，经精细结构分析和免疫细胞化学分析，选用人cDNA非放射性探针对其进行原位杂交检测。虽然用Southern杂交法用人cDNA探针检测了杂交，但在牙胚中未检测到mRNA的信号。此外，对人类淀粉原蛋白基因6外显子同源序列进行PCR扩增，在人类（400bp）、小鼠（400bp）、鸡（780bp）、凯门鳄（400bp）、乌龟（750bp）和蛇（750bp）的基因组DNA中均有扩增产物。但在蛙类、鱼类和环口动物中未获得相关产物。在对鸡、凯门鳄、龟和蛇的产品进行测序后，凯门鳄的DNA序列与人类淀粉原基因6外显子的对应率仅为48.5%。在鸡、龟和蛇的产品中发现了较低但轻微的同源性。因此，本研究表明，淀粉原蛋白基因样序列可能在脊椎动物中普遍保守。少