线粒体 DNA突变与心肌病关系的研究进展

人类某些疾病与线粒体DNA(mtDNA)基因组缺陷有关.本文就mtDNA突变与缺血性心肌病和肥厚型心肌病关系的研究加以回顾.目前的研究大多认为心肌缺血缺氧致氧化磷酸化紊乱,产生氧自由基损伤mtDNA,以及缺氧致氧化磷酸化过度诱导而损伤mtDNA,慢性损伤积累终致mtDNA片断缺失或点突变,主要表现出mtDNA5.0kb、7.4kb缺失及细胞色素b(cytb)基因上C15452A点突变;tRNA基因保守序列突变,致肌肉收缩蛋白合成缺陷,缺陷的收缩蛋白持续而无效的收缩可能会增加心肌对ATP的代谢需求,因此导致心肌肥厚.     

Use of subpopulation data in Australian forensic DNA casework

DNA profiling evidence presented in court should be accompanied by a reliable estimate of its evidential weight. In calculating such statistics, allele frequencies from commonly employed autosomal microsatellite loci are required. These allele frequencies should be collected at a level that appropriately represents the genetic diversity that exists in the population. Typically this occurs at broadly defined bio-geographic categories, such as Caucasian or Asian. Datasets are commonly administered at the jurisdictional level. This paper focuses on Australian jurisdictions and assesses whether this current practice is appropriate for Aboriginal Australian and Caucasian populations alike. In keeping with other studies we observe negligible differences between Caucasian populations within Australia when segregated geographically. However segregation of Aboriginal Australian population data along contemporary State and Territory lines appears to mask the diversity that exists within this subpopulation. For this reason datasets collated along more traditional lines may be more appropriate, particularly to distinguish the most genetically differentiated populations residing in the north of the continent.     

Successful DNA typing of ultrafiltered urines used to detect EPO doping

Endogenous and exogenous erythropoietin (EPO) present in urine can be distinguished according to their isoelectric profiles. This methodology requires urine samples to be concentrated about 200 to 1000 times with manipulations that should remove most of the cells occurring in the original sample. In this study, we tried to obtain DNA profiles from 10 ultrafiltered urines (retentates) in order to evaluate whether a formal genetic identification was technically feasible. No nuclear DNA profiles could be established from retentates, despite 34 PCR-cycles amplifications. Contrastingly, mitochondrial DNA (mtDNA) profiles were obtained for 9 out of the 10 retentates. Apart from some particularities, retentate mtDNA profiles were all distinct and matched mtDNA profiles of corresponding reference samples.     

Y-STR analysis on DNA mixture samples—Results of a collaborative project of the ENFSI DNA Working Group

The ENFSI (European Network of Forensic Science Institutes) DNA Working Group undertook a collaborative project on Y-STR typing of DNA mixture samples that were centrally prepared and thoroughly tested prior to the shipment. Four commercial Y-STR typing kits (Y-Filer, Applied Biosystems, Foster City, CA, USA; Argus Y Nonaplex, Biotype, Dresden, Germany; Powerplex Y, Promega, Madison, WI, USA; and DYSplex-3, SERAC, Bad Homburg, Germany) were used for the amplification of the mixture samples. The results of the study showed a striking inter-laboratory difference of kit performance as determined from the peak heights of the obtained Y-STR genotypes. Variation in quantity and quality of the shipped DNA can be excluded as reason for the observed differences because both samples and shipping conditions were found to be reproducible in an earlier study. The results suggest that in some cases a laboratory-specific optimization process is indicated to reach a comparable sensitivity for the analysis of minute amounts of DNA.     

Implementation of a robotized real-time PCR setup for the use of the Quantifiler™ Human DNA Quantification Kit

Human DNA quantification occupies a central role within the DNA analytic process of forensic casework samples as DNA quantification results have an important impact on the quality of the short tandem repeat data. Manual processing for the setup of quantification reactions can be time consuming and labor intensive. Therefore automation of quantitative real-time PCR setup was an important component of our DNA-analysis automation concept. Here we show the implementation of a robotized setup for the Quantifiler™ Human DNA Quantification Kit.     

Object-oriented Bayesian networks for paternity cases with allelic dependencies

This study extends the current use of Bayesian networks by incorporating the effects of allelic dependencies in paternity calculations. The use of object-oriented networks greatly simplify the process of building and interpreting forensic identification models, allowing researchers to solve new, more complex problems. We explore two paternity examples: the most common scenario where DNA evidence is available from the alleged father, the mother and the child; a more complex case where DNA is not available from the alleged father, but is available from the alleged father’s brother. Object-oriented networks are built, using HUGIN, for each example which incorporate the effects of allelic dependence caused by evolutionary relatedness.     

鸡传染性支气管炎病毒四川分离株N基因DNA免疫质粒的构建

采用SDS 蛋白酶K法抽提鸡传染性支气管炎病毒(IBV)四川分离株SAIBWJ的基因组RNA,参照基因库中的N基因序列设计了1对引物,扩增出了单一的DNA产物。将该产物插入pUC18载体的HindⅢ和BamHⅠ酶切位点,构建了pUC NWJ质粒。序列测定结果表明,获得的克隆质粒包含了全部N基因。再将N基因亚克隆到pcDNA3.1( )载体的HindⅢ和XhoⅠ酶切位点,构建了IBVSAIBWJN基因的DNA免疫质粒pcDNA NWJ。     

郑州地区食尸性苍蝇及幼虫mtDNA中COⅡ基因序列分析

目的 用mtDNA中细胞色素氧化酶辅酶Ⅱ(COⅡ)基因序列鉴定法医学中常见食尸性苍蝇及其幼虫的种类. 方法 收集郑州地区大白鼠尸体上的苍蝇及其幼虫,提取DNA,PCR扩增CO Ⅱ序列,测序,用Clustalx和MEGA 4.0软件对基因序列进行比对分析及构建系统进化树.结果 成虫与幼虫基因差异不明显,CO Ⅱ基因序列可以对棕尾别麻蝇、巨尾阿丽蝇和亮绿蝇进行鉴定,铜绿蝇与丝光绿蝇进化距离较近,CO Ⅱ序列不能将他们区分开,同时还发现巨尾阿丽蝇和亮绿蝇存在种群单核苷酸多态性. 结论 mtDNA中COⅡ序列能有效鉴定郑州地区部分常见食尸性苍蝇的种类,方法 简便、准确.     

改良高盐低pH方法提取陈旧大麻DNA初探

目的探讨建立室温保存10年队上大麻干叶及大麻树脂DNA提取方法。方法采用SDS及改良高盐低pH方法，改变提取缓冲液中β-巯基乙醇终浓度，增加用酚、氯仿快速抽提过程，提取新鲜和陈旧大麻（树脂）DNA，应用大麻叶绿体trnLintron引物进行PCR，琼脂糖凝胶电泳法检测扩增产物。结果用高盐低pH方法获得了10年以上大麻干叶及树脂清晰的电泳图谱，其中成功提取了1份23年陈旧大麻的DNA。结论高盐低pH方法操作简便、实用，可望用于陈旧、微量大麻植株的DNA检测，对于涉毒案件中特殊大麻标本的检验具有一定意义。     

利用AP-PCR对柔嫩艾美球虫不同地理株及其杂交株多态性的研究

用随机引物PCR(AP PCR)技术对鸡柔嫩艾美球虫(Eimeria tenella)黑龙江株(H)、山东株(S)及黑龙江株与山东株的杂交株(F1)进行了研究,同时对 2 个不同地理株的子代株与亲代株和F1 株的亲缘关系进行了分析。结果,鸡柔嫩艾美球虫不同地理株之间以及同一地理株的亲代与子代之间存在DNA多态性;并且不同地理株间的种内变异程度较大(Si=0.642 3~0.637 0);同一地理株亲代与子代之间的亲缘关系也稍有改变,但其变异程度不大(Si=0.983 0~0.981 1)。对F1 株的AP PCR分析发现,F1 株与H株、S株的相似值分别为0.692 9、0.682 5,介于H株与S株相似值和传代株间的相似值之间。结果表明,F 株既具有两亲本株的遗传性状,又发生了变异。