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Studies of molecular genetics on the basis of protein and nucleic acid preserved in fossil materials.

基于化石材料中保存的蛋白质和核酸的分子遗传学研究。

基本信息

批准号：
07454126
负责人：
CHIBA Satoshi
金额：
$ 5.06万
依托单位：
Shizuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07454126/
关键词：
Fossils,DNA Protein Molecular Biology Nucleotide sequences PCR technique Mollusca PCR 昆虫化石タンパク質化石DNA SDS‐PAGE

项目摘要

Condition of preservation of biomolecules such as protein and nucleic acid were examined, and potential of estimating phylongenetic relationship of fossil organisms by nsing these biomolecules was evaluated. Intracrystalline protein was extraced from four modern species (Patinopecten yessoensis, Umbonium giganteum, Sunetta menstrualis and Crassostrea gigas) and one fossil species (Patenipecten tokyoensis) of mollusca. Patenipecten tokyoensis were occurred from Pliocene deposits of Kakegawa fromation. Twenty-one species of protein were detected by electrophoretic analysis with SDS-PAGE.Protein molecules obtained from Patenipecten yessosensis and Patenipecten tokyoensis were restricted at the position of methionine. Restriction patterns of these molecules are coincident between modern species Patenipecten yessoensis and fossil species Patenipecten tokyoensis. Since these species are considered to be closely related, overall similarity of restriction patterns of protein between these spec … More ies suggest that protein have been preserved in the fossil materials.Phylogenetic relationship among the molluscan species estimated on the basis of the variations in the species of protein possessed by these molluscan species well corresponded to the phylogenetic relationships on the basis of nuclear DNA sequence (lrRNA). This suggests that the preserved protein in the fossils are useful to assess phylogenetic relationships. Nuclear and mitochondrial DNA were extracted from fossil insects (Donaciinae, Carabidae, Hydrophilinae) correceted from the Pleistocene and Holocene peat. These DNAs were amplified via PCR,and 300bp of nuclear 18SrRNA and mitochondrial 16SrRNA were sequenced. Sequences of the fossil species of Hydrophilinae were the closest to those of living species of Hydrophilinae among sequences obtained from Genbank. This suggests that DNAs of fossil Hydrophilinae species are amplified successfully and sequenced. This also suggests that DNA was preserved in the fossil insects occurred from peat. Less

考察了蛋白质、核酸等生物分子的保存条件，评价了利用这些生物分子评价化石生物系统发育关系的潜力。从软体动物的4个现代种(Patinopecten yessoensis、Umbonium Giganteum、Sunetta mentruis和Crassostrea gias)和1个化石种(Patenipecten Tokyoensis)中提取了晶内蛋白。Patenipecten tokyoensis产于上新世Kakegawa组沉积。用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法对21种蛋白质进行了分析，得到的蛋白质分子均限制在蛋氨酸的位置。这些分子的限制模式在现代物种Yessoens和化石物种Patenipecten tokyoensis之间是一致的。由于这些物种被认为是密切相关的，所以这些特定…之间蛋白质限制性内切酶图谱的总体相似性这些软体动物物种之间的系统发育关系由这些软体动物物种所拥有的蛋白质种类的差异来估计，与基于核DNA序列(LrRNA)的系统发育关系很好地对应。这表明化石中保存下来的蛋白质对评估系统发育关系是有用的。从更新世和全新世泥炭中提取了化石昆虫(石龙亚科、步甲亚科、水虫亚科)的核和线粒体DNA。通过聚合酶链式反应(PCR)扩增出这些DNA，并对其进行了测序。在Genbank获得的序列中，亲水亚科化石种的序列与活的亲水亚科物种的序列最接近。这表明成功地扩增出了嗜水虫化石的DNA并对其进行了测序。这也表明，从泥炭中产生的昆虫化石中保存了DNA。较少