全细胞PCR扩增用于鱼腥藻种类鉴定

传统鉴定藻种的方法主要是通过形态学观察的方法加以判断。蓝藻在自然条件和人工培养过程中 ,其形态、代谢能力等都可能发生变化 ,同时该过程需要的时间长 ,难以区分种或种以下的分类、单位 ,亦难以在水华暴发早期阶段准确鉴定。本文利用rDNA通用引物扩增 ,表明在 5 0 μL的反应体系中加入 2 0个鱼腥藻细胞能扩增出目的条带 ;对已知的鱼腥藻PC基因的分析设计引物 ,在BSA浓度为 0 2 %— 1% (w/v)下 ,全细胞扩增出实验室保存的四种鱼腥藻的部分PC以及PC IGS序列 ,序列分析结果表明PC I

鲤鱼3个亚种线粒体DNA ND5/6区段的PCR-RFLP分析及亚种间分子标记的确立

从鲤鱼3个亚种(Cyprinus carpio carpio,Cyprinus carpio haematopterus和Cyprinus carpiorubrofuscus)中选出具代表性的品系共10个,运用PCR方法,扩增出2.4 kb的mtDNA ND5/6区段.共用9种限制性内切酶进行酶切分析,结果表明,每个亚种拥有一种单倍型,有4种限制性内切酶(Dde I,Hae III,Taq I和Mbo I)酶切后产生了能将3个亚种区分开来的诊断性限制性酶切位点.可利用其作为鉴定3个亚种的遗传标记和遗传育种

RT-PCR法检测污染物甲基汞对调钙质基因表达的抑制

应用RT -PCR方法 ,通过用不同浓度的甲基汞对Hep2B细胞株进行不同时间长短的处理 ,检测其调钙质的mRNA表达水平 ,发现微量的甲基汞处理就可以引起细胞株细胞损伤 ,同时抑制调钙质mRNA的表达 ,进一步证实了在肝脏损伤的病理状态下调钙质基因的表达减少 ,从而为检测环境中甲基汞的含量提供了一个可能的分子生物学手段 .

原位杂交和原位PCR技术在鱼类基因定位中的应用

国家自然科学基金（项目编号：39730290）

水华蓝藻产毒特性的PCR检测法

特异引物对（ＴＯＸ　１Ｐ／１Ｆ；ＴＯＸ　２Ｐ／２Ｆ）用于检测微囊藻毒素合成酶基因ｍｃｙＢ片段在３８种水华蓝藻中的分布情况。结果显示，所有能产生微囊藻毒素的微囊藻都有特异扩增条带，非产毒株则没有。几种常规的毒性检测方法验证了ＰＣＲ方法所获结果的准确性。本研究发展了以全细胞ＰＣＲ法检测ｍｃｙＢ片断，说明全细胞ＰＣＲ检测法适用于不同来源的蓝藻材料。结果证明以ＤＮＡ为基础鉴别产毒和非产毒微囊藻及其他水华蓝藻的方法是可行和实用的。

集胞藻ORFs侧翼序列的PCR扩增和定向基因插入失活策略

针对集胞藻ＰＣＣ６８０３的１９２７个待定编码基因进行了两侧序列的ＰＣＲ扩增。４个亚株基因组在。ｓｌｌ０２６７－ｓｌｌ０２６８－ｓｌｌ０２６９区域的 ＰＣＲ扩增产物与 Ｋａｚｕｓａ ＤＮＡ数据存在差异。以叶绿素合成基因ｃｈｌＨ和ｃｈｌＬ为例，显示三片段连接ＰＣＲ产物可有效用于集胞藻６８０３基因组定向插入失活。

七株微囊藻系统进化关系的RAPD-PCR分析(英文)

应用RAPD－PCR的方法，选用24个随机引物，分析来自不同地区的7株微囊藻的基因组多态性．结果显示，Microcystis．viridis及M.wesenbergii明显与M．aeruginosa区分开．M．aeruginosa分为两个可视为不同种的异源分类单位．作为对照的Anabaenasp．7120与其他微囊藻株表现出完全不同的基因型及更远的遗传距离．此项研究表明，以基因型而不是表现型为基础，分析蓝藻种内及种间区别是可能的．因此，为解决蓝藻分类问题，特别是在种和属的水平上，提供了重要的线索．结合正在

七株微囊藻系统进化关系的RAPD-PCR分析

应用RAPD-PCR的方法,选用24个随机引物,分析来自不同地区的7株微囊藻的基因组多态性。结果显示,Microcystis.viridis及M.wesenbergii明显与M.aeruginosa区分开。M.aeruginosa分为两个可视为不同种的异源分类单位。作为对照的Anabaena sp.7120与其他微囊藻株表现出完全不同的基因型及更远的遗传距离。 此项研究表明,以基因型而不是表现型为基础,分析蓝藻种内及种间区别是可能的。因此,为解决蓝藻分类问题,特别是在种和属的水平上,提供了重要的线索。结合

Quantitative convergence analysis of multi-agent systems

We introduce a characterization of contraction for bounded convex sets. For discrete-time multi-agent systems we provide an explicit upperbound on the rate of convergence to a consensus under the assumptions of contractiveness and (weak) connectedness (across an interval.) Convergence is shown to be exponential when either the system or the function characterizing the contraction is linear. Copyright © 2007 IFAC.

PCR技术在环境生物学上的应用

ＰＣＲ技术在环境生物学上的应用邱东茹，吴振斌（中国科学院水生生物研究所武汉４３００７２）水是许多疾病的传播途径之一，为监测水环境、水处理系统和供水系统的卫生学质量，需要对水中的病原菌和病毒作检测，由于直接检查水中各种病原微生物方法复杂，如有些细菌很难...

PCR-DGGE Fingerprinting Analysis of Plankton Communities and Its Relationship to Lake Trophic Status

Plankton communities in eight lakes of different trophic status near Yangtze, China were characterized by using denatured gradient gel electrophoresis (DGGE). Various water quality parameters were also measured at each collection site. Following extraction of DNA from plankton communities, 16S rRNA and 18S rRNA genes were amplified with specific primers for prokaryotes and eukaryotes, respectively; DNA profiles were developed by DGGE. The plankton community of each lake had its own distinct DNA profile. The total number of bands identified at 34 sampling stations ranged from 37 to 111. Both prokaryotes and eukaryotes displayed complex fingerprints composed of a large number of bands: 16 to 59 bands were obtained with the prokaryotic primer set; 21 to 52 bands for the eukaryotic primer set. The DGGE-patterns were analyzed in relation to water quality parameters by canonical correspondence analysis (CCA). Temperature, pH, alkalinity, and the concentration of COD, TP and TN were strongly correlated with the DGGE patterns. The parameters that demonstrated a strong correlation to the DGGE fingerprints of the plankton community differed among lakes, suggesting that differences in the DGGE fingerprints were due mainly to lake trophic status. Results of the present study suggest that PCR-DGGE fingerprinting is an effective and precise method of identifying changes to plankton community composition, and therefore could be a useful ecological tool for monitoring the response of aquatic ecosystems to environmental perturbations.

Immunoglobulin joining (J) chain and its expression in the Chinese soft-shelled turtle (Pelodiscus sinensis)

The immunoglobulin (Ig) joining (J) chain plays an important role in the formation of polymeric Igs and their transport into secretions. In the present study, the cDNA sequence of J chain has been cloned from the Chinese soft-shelled turtle (Pelodiscus sinensis) by reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE). The cDNA sequence is 2347 bp in length and contains an open reading frame of 480 bp encoding 160 aa including the signal sequence. The deduced amino acid sequence has a high degree of homology with that of an already reported turtle J chain (80.7%), and of chicken (71.3%). By using real-time quantitative RT-PCR analysis, a significant up-regulation of J-chain transcripts was observed in spleen, kidney and blood of turtles injected with inactivated Aeromonas hydrophila, indicating the immune role of J chain in response to bacterial infection. (C) 2009 Elsevier B.V. All rights reserved.

Genomic organization and expression analysis of Toll-like receptor 3 in grass carp (Ctenopharyngodon idella)

Toll-like receptor 3 (TLR3) participates in the innate immune response by recognizing viral pathogens. To investigate grass carp immune system responding to GCRV (grass carp reovirus) infection, the full-length cDNA sequence and genomic organization of grass carp TLR3 (CiTLR3) was identified and characterized. The full-length genome sequence of CiTLR3 is composed of 5668 nucleotides, including five exons and four introns. The full-length of CiTLR3 cDNA is 3681 bp in length and encodes a polypeptide of 904 amino acids with an estimated molecular mass of 102,765 Da and a predicted isoelectric point of 8.35. Analysis of the deduced amino acid sequence indicated that CiTLR3 has four main structural domains, including a signal peptide sequence, 14 LRR (leucine-rich repeat) motifs, a transmembrane region and a TIR (Toll/interleukin-1 receptor) domain. It is most similar to the crucian carp (Carassius auratus) TLR3 amino acid sequence with an identity of 99%. Quantitative RT-PCR analysis showed that CiTLR3 transcripts were significantly up-regulated starting at day 1 and continued through day 7 following GCRV infection (P < 0.05). These data implied that CiTLR3 is involved in antiviral defense, provide molecular and functional information for grass carp TLR3, and implicate their role in mediating immune protection against grass carp viral diseases. (C) 2009 Elsevier Ltd. All rights reserved.