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在建立斑节对虾实验室养殖模式的基础上，对野外采集的有白斑综合症病毒（WSSV）病典型症状的斑节对虾初步纯化其病毒，并进行WSSV的PCR检测，得到WSSV的感染用样品。对健康斑节对虾分别进行浸浴感染，投喂感染和注射感染。对感染死亡个体进行WSSA的PCR检测和细菌检测，证实WSSV感染性和致死性。浸浴感染、投喂感染和注射感染的感染量分别为4mL/L、0.2g/10g虾体、1/2稀释液0.05mL/10g虾体，死亡开始时间分别为16d、42h、28h，三种感染模式最终死亡率100%，从开始死亡到全部死亡延续时间分别为15d、82h、44h。 相似文献
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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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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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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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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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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
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PCR法制备地高辛标记探针斑点杂交
检测白斑综合症病毒(WSSV) 总被引：2，自引：0，他引：2
用PCR法成功制备了DIG标记探针，探针长度为547bp，探针的产量为21.6ng/μL。此探针与随机引物合成探针检测样品灵敏度相近。用此探针核酸斑点杂交法检测了54尾中国对虾。结果表明此探针在对白斑综合症病毒的检测、对虾暴发性流行病的诊断等方面具有很高的应用价值。 相似文献