地质体中主要生物分子的研究方法及应用

现代分子生物学,生物化学及有机地球化学等领域的发展,为古生物研究者在发子水平上探索地质生物世界提供了新的概念和技术手段,本文着重讨论了可用于分子古生物学研究的生物分子,例如,维管植物的木质素等稳定生物聚合物,古生物细胞内的类脂化合物,碳水化合物,蛋白质及氨基酸和含遗传信息的核酸,分子古生物资料可用于探讨化石埋藏学,古生物分子系统学,生物演化模式与机制,分子演化速率,古生态环境再造等问题,本文较详细     

Preliminary Study on Applicability of Microsatellite DNA Primers from Parasite Protozoa Trypanosoma cruzi in Free-living Protozoa

1　Introduction　Generallyknownasacodominantgeneticmarker ,microsatellitehasbeenwidelyusedinstudiesonpopu lationgenetics,high resolutiongenotyping ,genemap ping ,evolution ,linkageanalysis ,conservationbiology ,behaviouralecology ,relationsbetweenparasite…     

自杀者的童年阴影

来自加拿大麦吉尔大学的一个研究小组发现，那些以自杀来结束生命的人的大脑与正常人的大脑有很大不同。即使“自杀者”大脑和“非自杀者”大脑在基因序列上差别不大，它们在“表观遗传标记”方面却存在明显不同。表观遗传学研究的是基因功能的改变，而这种改变并不涉及DNA序列的变化。遗传自父母的DNA，其序列在人的一生中不会改变，身体各部位的DNA序列相同。     

Species delimitation in the green algal genus Codium (Bryopsidales) from Korea using DNA barcoding

Codium, one of the largest marine green algal genera, is difficult to delimit species boundary accurately based on morphological identification only. DNA barcoding is a powerful tool for discriminating species of seaweeds. The plastid elongation factor TU(tuf A) is considered as maker to perform DNA barcoding of green algal species than rbc L gene due to universality and rapid evolution rate. We conducted DNA barcoding application to Codium specimens from the Jeju Island, Korea to overcome the limit of morphological identification and to confirm the species diversity. As a result of applying tuf A marker, we newly generated fifty-five tuf A barcodes to resolve eight species. Tuf A marker exhibited 6.1%–21.8% interspecific divergences, wider than the gap of rbc L exon 1,3.5%–11.5%. Molecular analysis of rbc L exon 1 sequences of Codium revealed eight distinct species like tuf A analysis separated in five phylogenetic groups. DNA barcoding of the genus Codium using tuf A marker is more helpful to overcome the limit of morphological identification, and this is more potential to reveal cryptic species and to resolve the relationships among subspecies than rbc L analysis alone. The complement of tuf A barcoding and rbc L analyses including morphology for the genus Codium in the northwestern Pacific will give much more reliable achievement for discovering species diversity and resolving the phylogenetic relationships.     

中国对虾选育群体“黄海1号”和野生群体不同组织基因组DNA甲基化的MSAP分析

DNA甲基化在调节动物生长发育和组织分化中发挥了重要作用。本研究主要从酶切、预扩和选扩等方面优化了中国对虾DNA甲基化分析的MSAP技术,给出了适合中国对虾MSAP分析的最近反应程序和体系,并采用该技术分别对中国对虾选育群体"黄海1号"和野生群体的肌肉、鳃和血细胞三种组织基因组DNA的CCGG甲基化水平进行分析。研究结果表明,中国对虾野生群体肌肉、鳃和血细胞的DNA甲基化比例分别为23.1%、22.3%和19.7%,选育群体"黄海1号"肌肉、鳃和血液的甲基化比例分别为21.4%、19.6%和18.9%。鳃组织的DNA甲基化水平在两群体中差异极显著(P <0.01),肌肉间的甲基化水平差异显著(P <0.05),而血细胞中甲基化水平差异不显著(P >0.05)。中国对虾野生群体和"黄海1号"同一组织间的甲基化水平不同(P ≤ 0.05),而不同组织间的甲基化水平也各不相同(P ≤ 0.05)。DNA甲基化多态性分析表明,野生群体和选育群体"黄海1号"的鳃和血细胞组织的甲基化多态性比例变化明显,而肌肉组织甲基化水平较稳定,这些变化趋势与CCGG位点的甲基化和去甲基化有关。     

重金属Cu、Pb对泥鳅(Misgurnus anguillicaudatus)卵细胞凋亡及DNA损伤的SCGE试验

采用室内暴露试验方法,以单细胞凝胶电泳技术(SCGE)检测,研究了Cu、Pb不同浓度梯度与不同暴露时间联合染毒对泥鳅卵细胞DNA的损伤.结果表明,各Cu、Pb浓度组DNA平均迁移长度增加,与阴性对照组比较差异有显著性(P＜0.05).此外,随着Cu、Pb染毒剂量的增加,各试验组DNA的平均迁移长度逐渐增加,在试验浓度梯度范围内(Cu 0.01mg/L+Pb 0.05mg/L、Cu 0.10mg/L+Pb 0.50mg/L、Cu 0.25mg/L+Pb 0.75mg/L),存在较为显著的剂量-效应关系(P＜0.05),但未见明显的时间-效应关系(P＞0.05).Cu、Pb可引起泥鳅卵细胞凋亡和DNA损伤,卵细胞的不同损伤水平可望作为较为理想的水环境基因毒性指标.     

基于cox1条形码的鱼肚DNA分子鉴定

为鉴定鱼肚的正品来源,运用一对特异性引物扩增并测定10 个不同价格（700～1 500 元/kg）鱼肚样本的线粒体DNA 细胞色素c 氧化酶Ⅰ（cox1）约660 bp 的序列.通过BLASTN 比对DNA 序列同源度,判定鱼肚所属种类分别为鲈形目石首鱼科（Sciaenidae）、马鲅科（Eleutheronema）、笛鲷科（Lutjanidae）、带鱼科（Trichiuridae）.cox1 条码序列获取便捷,基于此条码序列的物种鉴定技术可用于鱼肚种类鉴别.     

DNA Barcoding Assessment of Green Macroalgae inCoastal Zone Around Qingdao,China

An assessment with assistance of DNA barcoding was conducted on green macroalgae in coastal zone around Qingdao,China, during the period of April-December, 2011. Three markers were applied in molecular discrimination, including the plastidelongation factor tufA gene, the internal transcribed spacer （ITS） region of the ribosomal cistron and rubisco large subunit gene 3＇regions （rbcL-3P）. DNA barcoding discriminated 8 species, excluding species of genus Cladophora and Bryopsis due to failures inamplification. We ascertained and corrected 4 species identified by morphological methods for effectively assisting the classification.The gene tufA presented more advantages as an appropriate DNA marker with the strongest amplification success rate and speciesdiscrimination power than the other two genes. The poorest sequencing success largely handicapped the application of ITS. Samplesidentified by tufA and rbcL as Ulvaflexuosa were clustered into the clade of U. prolifera by ITS in the neighbor-joining tree. Confu-sion with discrimination of the complex of U. linza, U. procera and U. prolifera （as the LPP complex） still existed for the three DNAmarkers. Based on our results, rbcL is recommended as a preferred marker for assisting tufA to discriminate green macroalgae. Indistinguishing green-tide-forming Ulva species, the free-floating sample collected from the green tide in 2011 was proved to be iden-tical with U. prolifera in Yellow Sea for ITS and rbcL genes. This study presents a preliminary survey of green macroalgae distrib-uted in the coastal area around Qingdao, and proves that DNA barcoding is a powerful tool for taxonomy of green macroalgae.     

中部太平洋大眼金枪鱼(Thunnus obesus)种群遗传结构的初步研究

大眼金枪鱼(Thunnus obesus)是世界最重要的渔业资源之一,经济价值较高,捕捞压力的加大使中部太平洋大眼金枪鱼过度捕捞程度愈发严重。开展种群遗传结构研究将为大眼金枪鱼资源的可持续利用与开发提供科学依据。本文以取自中西太平洋和中东太平洋共182尾大眼金枪鱼个体为研究对象,分析研究其线粒体DNA控制区第一个高变区(the first hypervariable region,HVR-1)457bp的序列,共发现115个变异位点,定义了178种单倍型。序列多样性分析结果显示:8个采样点核苷酸多样性在0.03074—0.04362之间,单倍型多样性在各个采样点都较高,变动范围在0.996—1.000之间;分子方差分析显示99.97%的遗传变异性来自于种群内;群体间的高基因交流值Nm和低分化指数Fst揭示中东太平洋与中西太平洋群体间基因交流频繁,不存在显著的遗传分化。