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331.
C. Hernando M. Carrera I. Ribas N. Parear R. Baraibar J. Egocue C. Fuster 《黑龙江环境通报》2002,22(9):802-805
We describe three cases in which we used fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR) and comparative genomic hybridization (CGH) to characterize Y chromosome structural anomalies, unidentifiable by conventional G-banding. Case 1 was a 46,X,+mar karyotype; FISH analysis revealed an entire marker chromosome highlighted after hybridization with the Y chromosome painting probe. The PCR study showed the presence of Y chromosome markers AMG and SY620 and the absence of SY143, SY254 and SY147. CGH results confirmed the loss of Yq11.2-qter. These results indicated the presence of a deletion: del(Y)(q11.2). Case 2 was a 45,X [14]/46,XY[86] karyotype with a very small Y chromosome. The PCR study showed the presence of Y chromosome markers SY620 and AMG, and the absence of SY143, SY254 and SY147. CGH results showed gain of Yq11.2-pter and loss of Yq11.2-q12. These results show the presence of a Yp isodicentric: idic(Y)(q11.2). Case 3 was a 45,X,inv(9)(p11q12)[30]/46,X,idic(Y)(p11.3?),inv(9)(p11q12)[70] karyotype. The FISH signal covered all the abnormal Y chromosome using a Y chromosome paint. The PCR study showed the presence of Y chromosome markers AMG, SY620, SY143, SY254 and SY147. CGH only showed gain of Yq11.2-qter. These results support the presence of an unbalanced (Y;Y) translocation. Our results show that the combined use of molecular and classical cytogenetic methods in clinical diagnosis may allow a better delineation of the chromosome regions implicated in specific clinical disorders. Copyright © 2002 John Wiley & Sons, Ltd. 相似文献
332.
Most of cystic fibrosis (CF) pre-implantation genetic diagnosis (PGD) cases described to date are limited to the detection of ΔF508. Beside this predominant mutation, over 1000 mutations have been identified, rendering the development of a mutation-based PGD protocol impracticable. This is the reason why we, as well as the others, have developed PGD strategies on the basis of the identification of the pathogenic haplotype instead of the mutation(s). In a previous article, we reported the conditions for the co-amplification of two intragenic polymorphic markers and the F508 locus. Here we describe an improved protocol allowing the additional amplification of two new intragenic markers, intron 1 CA repeat (I1CA) and IVS17bTA. This new protocol should, theoretically, allow us to provide a diagnosis to all couples requiring PGD for CF. Using single lymphoblasts, we have tested four different PCR configurations, including one duplex, two triplexes and one quadruplex PCR. All of them gave results compatible with a clinical application. The number of single lymphoblasts tested in each series varied from 89 to 155. PCR efficiency ranged from 95.4 to 100%. A complete haplotype was achieved for 83.2 to 90.7% of the tested cells, with an allele drop out (ADO) rate comprised between 6.0 and 11.6%. We present here three cases that we performed either with the former test (one case using the triplex PCR combining F508, IVS8CA and IVS17bCA) or with the new one (one case using the triplex combining F508, I1CA and IVS17bTA and one case using a quadruplex test). We obtained two single pregnancies. Copyright © 2004 John Wiley & Sons, Ltd. 相似文献
333.
334.
分子生物学方法在微生物多样性及微生物生态研究中的应用 总被引:5,自引:1,他引:4
首先介绍了分子生物学技术在微生物多样性及微生物生态研究中应用的理论基础,以及在此基础上引入的分子生物学技术如:PCR、DNA杂交技术在分子层面上来研究微生物遗传多样性,并结合已有工作积累展望了分子微生物生态研究的发展前景.图2参20 相似文献
335.
Joyce C. Harper Dagan Wells Wirawit Piyamongkol Patrick Abou-Sleiman Angela Apessos Antonis Ioulianos Mary Davis Alpesh Doshi Paul Serhal Massimo Ranieri Charles Rodeck Joy D. A. Delhanty 《黑龙江环境通报》2002,22(6):525-533
We report our experience of 14 preimplantation genetic diagnosis (PGD) cycles in eight couples carrying five different single gene disorders, during the last 18 months. Diagnoses were performed for myotonic dystrophy (DM), cystic fibrosis (CF) [ΔF508 and exon 4 (621+1 G>T)], fragile X and CF simultaneously, and two disorders for which PGD had not been previously attempted, namely neurofibromatosis type 2 (NF2) and Crouzon syndrome. Diagnoses for single gene disorders were carried out on ideally two blastomeres biopsied from Day 3 embryos. A highly polymorphic marker was included in each diagnosis to control against contamination. For the dominant disorders, where possible, linked polymorphisms provided an additional means of determining the genotype of the embryo hence reducing the risk of misdiagnosis due to allele dropout (ADO). Multiplex fluorescent polymerase chain reaction (F-PCR) was used in all cases, followed by fragment analysis and/or single-stranded conformation polymorphism (SSCP) for genotyping. Embryo transfer was performed in 13 cycles resulting in one biochemical pregnancy for CF, three normal deliveries (a twin and a singleton) and one early miscarriage for DM and a singleton for Crouzon syndrome. In each case the untransferred embryos were used to confirm the diagnoses performed on the biopsied cells. The results were concordant in all cases. The inclusion of a polymorphic marker allowed the detection of extraneous DNA contamination in two cells from one case. Knowing the genotype of the contaminating DNA allowed its origin to be traced. All five pregnancies were obtained from embryos in which two blastomeres were biopsied for the diagnosis. Our data demonstrate the successful strategy of using multiplex PCR to simultaneously amplify the mutation site and a polymorphic locus, fluorescent PCR technology to achieve greater sensitivity, and two-cell biopsy to increase the efficiency and success of diagnoses. Copyright © 2002 John Wiley & Sons, Ltd. 相似文献
336.
Xunwen ChenHui Li Wai Fung ChanChuan Wu Fuyong WuShengchun Wu Ming Hung Wong 《Chemosphere》2012,89(10):1248-1254
As a silicon hyperaccumulator, lowland rice takes up higher levels of As than many other plants due to silicic acid and arsenite sharing the same transporters (Lsi1 and Lsi2). Glomus intraradices (AH01) was inoculated to rice under different arsenite concentrations (0, 2 and 8 μM) in order to investigate the interactions between arbuscular mycorrhizal fungus and rice on the accumulation of arsenite. The relative mRNA expressions of Lsi1 and Lsi2 resulted in a down-regulating trend in mycorrhizal plants. Under 2 μM arsenite treatments, Lsi1 and Lsi2 were significantly decreased, by 0.7-fold (P < 0.05) and 0.5-fold (P < 0.01), respectively, in mycorrhizal plants when compared with non-mycorrhizal plants. This led to the decrease of arsenite uptake per unit of root dry mass. No organic As species were detected in both roots and shoots. The As(III)/As(V) ratios indicated that mycorrhizal plants immobilized most of the arsenite proportion in the roots and prevented its translocation from the roots to the shoots. 相似文献
337.
江苏省代表性水源地抗生素及抗性基因赋存现状 总被引:6,自引:5,他引:1
抗生素和抗生素抗性基因(antibiotic resistance genes,ARGs)是环境中重要的新兴污染物,为探明江苏省代表性水源地多种环境介质中抗生素和ARGs的污染水平及影响因素,于2018年12月和2019年6月采集苏北、苏中和苏南的5处代表性集中式饮用水水源地取水口处水体、表层沉积物和石相附着生物膜样品,对3种介质中10种代表性抗生素浓度、1类整合子酶基因intl1和7种代表性ARGs的绝对丰度进行检测分析.结果表明,5处水源地中目标抗生素和ARGs处于较低赋存水平.磺胺类抗生素在水体、表层沉积物和附着生物膜中的赋存量分别为NF(未检出)~37.4 ng·L-1,NF~47.3 ng·g-1和NF~3759.1 ng·g-1.喹诺酮类抗生素在3种介质中的浓度和含量分别为NF~5.3 ng·L-1、0.4~32.5 ng·g-1和NF~4220.9 ng·g-1.目的ARGs中,sul1、sul2、tetW和tetQ的检出率为100%,其中磺胺类抗生素ARGs,即sul1和sul2基因丰度最高.表层沉积物和附着生物膜中的ARGs丰度相当,高于水体中ARGs的丰度.网络分析结果表明,所属拟杆菌门、变形菌门、厚壁菌门、疣微菌门和放线菌门的细菌最有可能成为代表性水源地中ARGs的潜在宿主,在ARGs的扩散和转移过程中起重要作用.研究结果对于江苏省集中式饮用水源地水环境质量状况评估和水质安全保障具有一定的科学指导意义. 相似文献
338.
基于全球定位系统(GPS)的电动车辆实时监控系统设计 总被引:1,自引:0,他引:1
电动汽车以其良好的环保效应和灵活多样的能源利用形式,已成为国际社会普遍公认的汽车工业的重要发展方向之一,作为凸显可持续发展理念和国家科技产业实力的窗口,电动汽车的规模化应用将成为彰显2008北京“绿色奥运、科技奥运”主题的鲜明标志。为确保电动汽车奥运应用的安全性与可靠性,笔者提出了基于GPS的电动车辆实时监控系统的总体设计。该系统由车载信息采集终端(OBICT)、车辆运行监控(VOP)和无线通讯网络(WCN)3个子系统构成,并能对电动汽车运行中的电池及关键部件工作状态、车辆位置等数据进行实时采集与传输。在给出系统总体结构设计的基础上,对该车载信息采集终端、车辆运行监控子系统的数据流程进行分析,并提出了相应的软/硬件架构技术、设计方法以及实现的功能。 相似文献
339.
采用PCR-DGGE等分子生物学技术。从平行AN/AO工艺六个反应池的活性污泥样品中提取总DNA,采用套式PCR(nested PCR)进行两轮PCR扩增。对α-Proteobaeteria、β-Proteobacteria的16S rDNA片段进行DGGE(变性梯度凝胶电泳)分离,从而分析活性污泥样品中该类除磷菌的种群结构。结合测序技术并通过NCBI(美国国立生物技术信息中心)基因库比对,证实生物除磷过程中存在着丰富的α-Proteobacteria、β-Proteobacteria,同时可以得到其中优势类群的初步认识。结果表明,PCR-DGGE结舍测序技术可以用于污水处理过程中样品的除磷菌群落结构分析。 相似文献
340.
在化工企业中通过系统科学辨识危险源,针对危险源安全控制数据,采用OPC通讯接口标准,利用网关机和防火墙隔离技术,实现安全的与生产系统数据链接,建成危险源实时监控与应急预警信息系统的有效链接;通过Web实时画面对现场危险源数据进行监控,数据出现波动立即启动相应预案中的应急措施,利用移动公司的企信通服务,利用面向对象语言采用Socket接口编程,自主开发短信模块接口程序;通过系统调用预警模块实现短信监报功能,应用短信瞬间发布应急预警指令,为应急处理赢得时间,有效地遏制安全生产事故的发生。 相似文献