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聚合酶链式反应是现代生物学领域一种非常重要的技术，本文介绍了这种技术在对虾病毒研究中的具体应用，包括病原检测、流行病学调查、寄生部位确定、同源性分析、DNA多态性分析、基因克隆与探针制备等。 相似文献
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The fragments of 350 bp in 28S rRNA from the closely related monogenea of trematoda, Neobenedenia girellae and N. melleni are obtained by polymerase chain reaction (PCR) amplified using a couple of special primers and then sequenced. The results show that the comparison of 28S rRNA sequences, with only a base varying in 337 bp accounting for 0.3% genetic difference, from the relative species N. girellae and N. melleni parasitized on the different fishes in different farms displays that they possess a very high genetic similarity of 99.7%, higher than that of 99.41% for the single species N. melleni sampled in different areas, and the intraspecific divergence of N.melleni is 0.59%. Meanwhile, the interspecific differences between the two Neobenedenia and three Benedenia (i.e., B. lutjani, B. rohdei and B. seriolae) range from 2.08% tol 1.73%. In addition, UPGMA and MP molecular phylogenetic trees are constructed and proved to be consistent with each other. Though the morphological characteristics and the results of genetic diversity for the two Neobenedenia show a high similarity, whether they belong to a single species or not are still undefined, and the more genes of them should be further investigated, in combination with the systematical and detailed morphological study. 相似文献
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介绍了一种制备海洋浮游生物PCR模板DNA的方法——碱煮法。取少量海水样品(40μL)．直接经0．25mol／L艺机NaOH和99℃温育处理裂解细胞,从而得到环境样品总DNA。实验证明,该方法制备的模板DNA可用于细菌核糖体RNA、浮游植物叶绿体rbcL和浮游动物线粒体COI等基因的PCR扩增。但是,在使用通用引物扩增细菌基因时,假阳性很难避免。该法制备的模板DNA适用于扩增非细菌基因或者特异引物界定的细菌基因。该方法较传统制备方法快速、简便,样品需要量减少,适用于浮游生物分子遗传多样性研究。 相似文献
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对含牛粪、玉米秸秆、废菜叶和废纸的农村垃圾进行堆肥实验,实验分为预堆腐和蚯蚓处理2个阶段。预堆腐以EM(Effective Microorganisms)原液和有机物料腐熟剂(FU)为促腐剂,预堆腐结束后,堆料继续进行蚯蚓处理。利用聚合酶链反应与变性梯度凝胶电泳联用技术(PCR-DGGE)分析了对照组(CK)及适合蚯蚓生长繁殖的4‰FU和5‰EM组蚯蚓处理前后细菌和真菌群落中的优势菌群。结果表明:在4‰FU组、5‰EM组和CK组的细菌群落中均存在且占有优势的种群与藤黄单胞菌属和梭状芽孢杆菌属具有较高的亲缘性;在5‰EM组出现的占优势的真菌种群与轮枝孢属具有较高的亲缘性,在CK组中出现的占优势的真菌种群与镰孢菌属和曲霉属具有较高的亲缘性。 相似文献
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《台湾海峡》2011,(4):595-596
论文与报告基于连续沥取的富钴结壳成凶机制的探讨………………………………………………………………………………蔡毅华,黄奕普,邢娜（1）饥饿对斜带石斑鱼稚鱼胃肠道内分泌细胞的形态、定位和分布密度的影响…………………………………………………翁幼竹,方永强（10）地芽孢杆菌属DYth03DNA聚合酶基因的克隆、表达及性质分析……………………………………………………罗淑娅,李侃,徐丽美（17）杂色鲍血蓝蛋白HtHI基因第16内含子SNP密度的分析…………………………………………梁翠,陈军,游伟伟,王德祥,柯才焕（21）大亚湾大鹏澳海域表层沉积物中浮游植物休眠体集群的季节变化…………………………………………付永虎,王朝晖,康伟,杨宇峰（27） 相似文献
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We isolated 4 Norwalk-like viruses (NLVs) contaminated oysters from 33 Chinese oysters collected from local commer-cial sources of Shandong Province. After amplification of the RNA-dependent RNA polymerase (RdRp) region of NLVs genomes with RT-PCR, the open reading frame 1 (ORF1) of the RdRp was sequenced and subjected to multiple-sequence alignment. The re-suits showed that NLVs in the four isolates belong to genogroup Ⅱ. The sequence comparison showed that the similarity between four Chinese oyster isolates were higher than 99.0%, which indicated that NLVs prevalent in close areas have high homogeneity in genome sequences. In addition, the most conserved sequences between diverse NLVs were used to design primers and TaqMan probes, then the real-time quantitative PCR assay was performed. According to the standard curve of GII NLVs, the original amounts (copies) of NLVs in positive patient's fecal isolate, positive Japanese oyster isolate, and the Chinese oyster isolate were 8.9×108, 1.25×108 and 4.7×101 respectively. The detecting limit of NLVs was 1×101 copies. This study will be helpful for routine diagnosis of NLVs pathogens in foods and thus for avoiding food poisoning in the future. 相似文献
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斑节对虾虾苗白斑综合症杆状病毒的检测和养殖跟踪 总被引：1，自引：0，他引：1
利用２步ＰＣＲ检测技术对湛江和阳西地区的虾苗进行白斑综合病杆状病毒（ＷＳＳＶ）检测。在仔虾２日龄最早检测得到病毒,有２５％的虾苗带有ＷＳＳＶ病毒。带病毒虾苗和不带病毒虾苗分别在不同养殖模式的养殖过程中跟踪。前者在湛江湖光镇普通虾塘跟踪养殖,它们在变化的环境中容易发病,ｐＨ、盐度和温度是重要的诱发因子,在养殖５０～６０ｄ时发病死亡;后者在高位池和普通池养殖跟踪,它们对变化的环境有较大的适应性,养殖时间为８０～１１０ｄ,在相对优良的养殖技术条件下大部分可望养殖成功。环境中有ＷＳＳＶ病原传入,不带病毒虾苗在养殖后期可以带有ＷＳＳＶ病毒,出现白斑虾。跟踪有普通池有爆发病害,但是时间延后,跟踪的高位池没有爆发病害。 相似文献
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参照鱼类病毒性出血性败血症病毒序列,根据PCR引物设计的原则,设计巢式PCR引物,采用巢式逆转录聚合酶链反应(RT-PCR)方法,对如何快速检测鱼类病毒性出血性败血症病的方法进行了较为系统的研究,并对RT-PCR的反应条件进行了优化.结果表明,巢式RT-PCR扩增获得279bp的特异性片断,阴性对照无扩增条带.巢式RT-PCR扩增出的特异性片段经测序分析,结果证实与报道的序列完全一致.该方法最低可检测出0.1pg的鱼类病毒性出血性败血症病毒RNA.初步建立了VHSV的巢式RT-PCR检测方法,该方法灵敏、特异,可为VHSV的检测提供一个快速、有效的手段. 相似文献
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为了获取具有广泛适用性的DNA聚合酶,将深海热液区获得的超嗜热古菌Palaeococcus pacificus DY20341的Pol B DNA聚合酶和d UTPase进行原核表达和纯化,将得到的Pol B DNA聚合酶及d UTPase混合后用于扩增古菌、细菌和真核生物的基因.通过PCR反应成功地扩增了商业化Pfu DNA聚合酶所不能扩增的Palaeococcus pacificus DY2034的琼脂酶基因.由此可知,尽管商业化的DNA聚合酶多种多样,针对不同扩增目的片段所需的特异DNA聚合酶体系仍然是必要的,这是又一嗜热古菌B家族DNA聚合酶应用的报道. 相似文献