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31.
In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress. 相似文献
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于1997年6月在青岛市沙子口海域采集文昌鱼样品，采用RT－PCR方法进行青岛文昌鱼LIM类同源框基因片段的扩增研究。结果表明，用一对引物一次PCR扩增的方法未能得到特异的PCR产物，用一对引物两次PCR扩增的方法也同样未果。在对套式PCR进行研究时发现，用普遍PCR方法进行的套式PCR结果不够理想，而用三步两段法和四步两段法套式PCR则可进行成功的扩增。用这两种方法扩增的PCR产物具有高度特异性。相比之下，用简并的引物对未知的基因片段进行PCR扩增时，三步两段法和四步两段法的套式PCR应是首选方法。 相似文献
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采用聚合酶链式反应限制性片段长度多态性分析(PCR-RFLP)技术对劳盆地热液区TVG9站位沉积物中细菌、古菌多样性进行了调查,并构建其细菌、古菌种群的16S rRNA基因文库.研究结果表明,该站位细菌包含变形杆菌(Proteobacteria)、硝化螺旋菌(Nitrospira),疣微菌(Verrucomicrobia)等3个类群,其中变形杆菌占据优势地位,由Alphaproteobactria,Gamaproteobactria,Deltaproteobactria等3个亚群组成.古菌包含泉古菌(Crenarchaeota)和广古菌(Euryarchaeota),其中泉古菌占优势,由MG Ⅰ (Marine Group Ⅰ)和MCG(Miscellaneous Crenarchaeotal Group)两类群组成,而广古菌主要由MBGE(Marine Benthic Group E)组成.旨在揭示深海热液区的微生物多样性,为生态环境的研究提供理论支持. 相似文献
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为了解色素控制基因与壳色的关系,本实验用荧光定量聚合酶链式反应(Polymerase Chain Reaction,PCR)技术分析了酪氨酸酶基因(Tyrosinase,TYR)在菲律宾蛤仔(Ruditapes philippinarum)4种壳色蛤仔和7种组织中的表达特性。结果表明,酪氨酸酶基因在鳃、外套膜、闭壳肌、性腺、内脏团、水管和唇瓣中均有表达,其中外套膜中表达量较高,其次为鳃,在闭壳肌中表达量最少。不同的酪氨酸酶基因在4种壳色的表达量不同,酪氨酸酶基因TYR2、TYR3、TYR6和TYR9在白斑马蛤中表达量居高,且与斑马蛤和白蛤差异显著(P0.05)。TYR2、TYR6和TYR10在橙蛤中表达量最高,推测与橙色形成有关。TYR11在斑马蛤中表达量最高,且与白蛤和白斑马蛤差异显著(P0.05),推测与背景色形成有关。系统发育分析结果表明,TYR3与马氏珠母贝(Pinctada martensii)同源性最高为64%。TYR10与加州双斑蛸(Octopus bimaculoides)同源性最高,为53%。与长牡蛎进化关系最近。 相似文献
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长棘海星(Acanthaster planci)的暴发是导致我国南海乃至印度—太平洋海域珊瑚礁退化的主要原因之一。浮游幼体的密度是决定成体种群是否暴发的重要指标,但是由于幼体肉眼不可见且不易分辨,常规调查和显微镜观察均无法有效检测到自然海域的长棘海星幼体,因此迫切需要开发一种高灵敏性且特异性的长棘海星幼体检测技术。本研究针对长棘海星幼体线粒体细胞色素氧化酶亚基I(mt COI,mitochondrial cytochrome oxidase subunit I)基因序列,建立了基于聚合酶链式反应(PCR,polymerase chain reaction)的长棘海星幼体特异性检测技术,并对西沙七连屿珊瑚礁海域的长棘海星幼体进行了检测。结果表明,设计筛选的4对特异性引物均可以扩增长棘海星Apmt COI基因片段,且与蓝指海星(Linckia laevigata)、面包海星(Culcita novaeguineae)、粒皮海星(Choriaster granulatus)和吕宋棘海星(Echinaster luzonicus)没有交叉反应。在退火温度为58.5℃时,引物2aooni F/2... 相似文献
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弧菌(Vibrio)是各种海洋环境的正常细菌区系的重要组成成分,也是水生动物与人类的重要病原之一。近年来出现了各种灵敏、快速、自动化且易于操作的分子分型方法,它们在弧菌分型中的应用,对揭示弧菌菌株在分子水平上的变异和进化规律、寻找病原及调查传播途径、控制疾病暴发有至关重要的作用。应用于弧菌的各种分子分型方法可分为依赖于PCR分型技术和不依赖于PCR分型技术,各种分型技术具有各自的优、缺点;其中前者由于高度的特异性、敏感性、可操作性、准确性、快速和费用低廉而得到广泛的应用,但重复性较差;后者由于基于生物体全基因组的操作,所以相对于依赖于PCR技术的分型方法结果更为可信,缺点是操作繁琐,对某些特殊细菌不适用。现今的国内外弧菌分型中公认的"金标准"的PFGE方法与近年来新发展起来的IRS-PCR分型方法结合,同时利用现有的数据建立国际分型数据库,实现信息和技术标准与共享,是弧菌分型的较好方法。 相似文献
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Cloning and Analysis of Calmodulin Gene from Porphyra yezoensis Ueda （Bangiales, Rhodophyta） 总被引：2，自引：0，他引：2
In order to understand the mechanisms of signal transduction and anti-desiccation mechanisms of Porphyra yezoensis, cDNA and its genomic sequence of Calmodulin gene (CaM) was cloned by the technique of polymerase chain reaction (PCR) based
on the analysis of P. yezoensis ESTs from dbEST database. The result shows that the full-length cDNA of CaM consists of 603 bps including an ORF encoding
for 151 amino acids and a terminate codon UGA, while the length of genomic sequence is 1231 bps including 2 exons and 1 intron.
The average GC content of the coding region is 58.77%, while the GC content of the third position of this gene is as high
as 82.23%. Four Ca2+ binding sites (EF-hand) are found in this gene. The predicted molecular mass of the deduced peptide is 16688.72 Da and the
pI is 4.222. By aligning with known CaM genes, the similarity of CaM gene sequence with homologous genes in Chlamydomonas incerta and Chlamydomonas reinhardtii is 72.7% and 72.2% respectively, and the similarity of the deduced amino acid sequence of CaM gene with homologous genes
in C. incerta and C. reinhardtii are both 71.5%. This is the first report on CaM from a species of Rhodophyta. 相似文献
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用逆转录聚合酶链反应检测草鱼出血病病毒的研究 总被引：3，自引：1，他引：3
于１９９４年５月－１９９５年７月，根据已克隆的草鱼出现血病病毒ＧＣＨＶ－８６１株ｃＤＮＡ的部分序列，设计合成了两对ＰＣＲ引物，采用ＲＴ－ＰＣＲ技术对ＧＣＨＶ－８６１ＧＣＨＶ－８７３两病毒株的ｄｓＲＮＡ进行扩增，结果表明，两对引物仅能特异地检测了ＧＣＨＶ－８６１病毒株核素的存在，而不能对ＧＣＨＶ－８７３病毒株的核酸进行特异扩增，该方法是最小可检测出０．１ｐｇ纯化的ＧＣＨＶ－８６１病毒ｄｓＲＮＡ，采用 相似文献