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[目的]探索并建立悬铃木方翅网蝽总DNA提取方法。[方法]采用改良SDS法结合Silica吸附柱提取悬铃木方翅网蝽总DNA,通过分光光度法、琼脂糖凝胶电泳对获得的DNA进行浓度、纯度及质量检测。利用PCR技术,分别扩增悬铃木方翅网蝽线粒体基因细胞色素氧化酶亚基Ⅰ(COⅠ)和基因组28S基因,克隆测序。[结果]悬铃木方翅网蝽总DNA浓度为52.8μg/ml,A260/A280为2.06,琼脂糖凝胶电泳条带清晰。PCR扩增产物条带清晰特异,经克隆并测序后表明为悬铃木方翅网蝽COⅠ和28S基因。[结论]该方法可得到较纯净完整和扩增效果良好的DNA,能满足进一步分子生物学研究的需要。 相似文献
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Cryptocaryon irritans recombinant proteins as potential antigens for sero‐surveillance of cryptocaryonosis 下载免费PDF全文
Cryptocaryonosis is a major problem for mariculture, and the absence of suitable sero‐surveillance tools for the detection of cryptocaryonosis makes it difficult to screen Cryptocaryon irritans‐infected fish, particularly asymptomatic fish. In this study, we proposed a serum‐based assay using selected C. irritans proteins to screen infected and asymptomatic fish. Eight highly expressed genes were chosen from an earlier study on C. irritans expressed sequence tags and ciliate glutamine codons were converted to universal glutamine codons. The chemically synthesized C. irritans genes were then expressed in an Escherichia coli expression host under optimized conditions. Five C. irritans proteins were successfully expressed in E. coli and purified by affinity chromatography. These proteins were used as antigens in an enzyme‐linked immunosorbent assay (ELISA) to screen sera from experimentally immunized fish and naturally infected fish. Sera from both categories of fish reacted equally well with the expressed C. irritans recombinant proteins as well as with sonicated theronts. This study demonstrated the utility of producing ciliate recombinant proteins in a heterologous expression host. An ELISA was successfully developed to diagnose infected and asymptomatic fish using the recombinant proteins as antigens. 相似文献
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Parasitic infections caused by Tetrahymena sp. constitute a serious problem in guppies, Poecilia reticulata. Tetrahymena was isolated from skin lesions of naturally infected guppies in a commercial aquaculture farm, cultured in vitro and used in subsequent experimental infections. In addition to guppies, angelfish, Pterophyllum scalare, platyfish, Xiphophorus maculates, and neontetra, Paracheirodon innesi, were susceptible, whereas tilapia (Oreochromis niloticus xO. aureus) was resistant. The ciliate had a high affinity for dead fish. Skin abrasion did not affect the infection, but fish with gas bubble disease exhibited a significantly higher infection than non-affected fish. Infection was significantly higher when fish were exposed to high levels of ammonia, high organic load and low water temperatures. Under shipment conditions, infection was significantly elevated. Full recovery was achieved at a low fish density. Results suggest that poor environmental and physiological conditions enhance infection with Tetrahymena sp. 相似文献
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SUMMARY: Experimental infection of Tetrahymena pyriformis was conducted on ornamental fishes with skin wounded by acetic acid treatment. Among fishes used in this experimental infection, guppy Poecilia reticulata , pristella Pristella maxillaris , neontetra Paracheirodon innesi and cherry barbs Puntius titteya proved to be sensitive to challenge, while medaka Oryzias latipes , dwarf gourami Colisa lalia , goldfish Carassius auratus , platy Xiphophorus maculatus and angelfish Pterophyllum scalare were resistant. In catfish Corydoras aeneus infection was unsuccessful. Exposure to Tetrahymena at ≥100 cells/mL at 25 to 30°C and at pH of 6.0 to 8.0 resulted in the successful infection of some sensitive fish species. Histological and bacteriological observations suggest that deeply destroyed skin tissues from the acid-treated method are the primary factors in successful infection. 相似文献
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应用透射电镜术显示了腹毛目纤毛虫新伪尾柱虫(Pseudourostyla nova)的皮层表膜及表膜下细胞质、皮层微管胞器和其他细胞质胞器。结果显示:新伪尾柱虫纤毛器基部附属微管较为短小,细胞表膜下成束排列的微管结构与进化程度较高的纤毛虫的微管胞器表现出诸多差异,认为腹毛目纤毛虫棘毛基部微管的发达程度是与其各类群细胞的分化程度密切联系的。 相似文献
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采用温室土培盆栽试验和水培试验方法,研究了重金属复合污染条件下香薷的Cu、Zn、Cd、Pb耐性和吸收性.结果表明,土培试验中,各重金属处理条件未对香薷造成毒害,香薷均生长良好,且各处理间生物量无显著差异;香薷地上部Cu、Zn、Cd、Pb浓度最高分别为12.1、100、0.83、11.3 mg/kg.水培试验条件下,Cu 50 μmol/L、Zn 200 μmol/L+Cd 10 μmol/L及Pb 100 μmol/L处理时香薷均生长良好,且生物量与对照均无显著差异,根部Cu、Zn、Cd、Pb浓度最高为2 270、1 648、1 029、25 331 mg/kg; Cu 100 μmol/L处理时香薷生物量显著低于对照,但并未死亡,该处理下香薷叶片丙二醛、还原型谷胱甘肽、可溶性糖浓度均显著高于对照.两组试验表明,香薷对Cu、Zn、Cd、Pb有极强的耐性,是一种良好的可栽种于重金属污染土壤的经济作物. 相似文献
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