口蹄疫病毒VP1植物表达载体的构建

P1 T重组质粒上含有口蹄疫病毒 (FMDV)GD10分离株的p1cDNA片段 ,以此为模板 ,用PCR方法扩增其中的VP1基因 ,获得大小约 6 40bp的片段。该片段用BglⅡ和BstEⅡ酶切消化后克隆至表达载体 pCAMBIA130 5 .2 ,转化EcoliTOP10感受态细胞。重组质粒经PCR、酶切及序列分析 ,证实VP1基因处于CaMV35S启动子控制 ,且读码框正确     

冰川表面冰尘洞内病毒研究进展

病毒在驱动生态系统高生物多样性形成以及生态系统营养物和有机物的循环过程中均发挥着重要的作用。冰川洞作为冰冻圈微生物活动的热点, 为病毒提供了特殊的生存环境, 驱使病毒产生了生存与适应机制。通过综述国际上近些年来关于冰川表面冰尘洞内病毒的主要研究成果, 内容涉及冰尘洞在冰川生态系统中的重要性, 冰尘洞内病毒的丰度与生产力, 冰尘洞内病毒的遗传多样性, 病毒与宿主的相互作用关系, 以及病毒对于冰尘洞碳循环和食物网的重要性。此外, 还总结了当前冰尘洞内病毒研究存在的空白： 形态多样性、 降解率、 分离培养、 冰川表面散落冰尘内病毒, 指出了未来研究需要重点关注的几个方面, 为进一步研究病毒对于冰尘洞生态系统的重要影响提供科学参考, 并且对青藏高原冰川表面冰尘洞内病毒未来的研究提出了展望。     

PCR检测对虾白斑综合征病毒(WSSV)中假阴性的探讨

采用PCR、核酸探针斑点杂交、组织切片等方法研究了PCR检测白斑综合征病毒(WSSV)中的假阴性问题。设计了一对引物用于PCR检测WSSV,PCR扩增的产物长度为235bp,检出极限为0.1pg WSSV DNA。结果表明,PCR检测15例攻毒螯虾鳃样品时出现一例阴性,而经10—106倍稀释后的样品却呈现阳性,推断PCR出现了假阴性。核酸探针斑点杂交及组织切片结果表明注射WSSV的螯虾鳃组织确已被严重感染,进一步证实了PCR假阴性。根据PCR的检出极限及模板稀释梯度,推算出该PCR能成功扩增的引物与模板浓度的比例范围约为2.4×105—2.4×1010。     

Reduced removal of bacteriophage MS2 in during basin infiltration managed aquifer recharge as basin sand is exposed to infiltration water

Basin infiltration managed aquifer recharge (MAR) is a commonly used method for storing and treating surface water to be used as drinking water. This study examined how the removal of bacteriophage MS2 was affected by the relative age of the sand used for basin infiltration MAR at 4 °C using batch experiments (static and agitated) and column experiments. The sand and the water used in all experiments were characteristic of that used at a basin infiltration MAR scheme in Uppsala, Sweden. Experimental data was fit with numerical models describing the fate and transport of virus in soil—water systems. The “used” sand that had been subjected to intermittent infiltration over a period of 8 years had 15 times the amount of organic carbon than the “new” sand, which had yet to be used for infiltration. Results showed that attachment of MS2 to the new sand in batch experiments was relatively irreversible. Attachment to the used sand was reversible. Inactivation of MS2 was slowed when it was attached to the used sand. Results for the column experiments showed that the removal rate of MS2 was significantly lower in columns of used sand than in columns of new sand. Simulations indicated that MS2 would be entirely removed in the infiltration basins with new sand. Less than 3 log removals was estimated for basins with used sand. Reduced removal of MS2 by the used sand was deemed to be most likely due to organic coatings on the used sand. Results of this study give deeper insight into the mechanisms responsible for removing virus in infiltration basins and how those mechanisms will change as the sand in the basin is exposed to infiltration water.     

White spot syndrome virus inactivation study by using gamma irradiation

The present study was conducted to investigate the effect of gamma irradiation on white spot syndrome virus(WSSV). White spot syndrome virus is a pathogen of major economic importance in cultured penaeid shrimp industries. White spot disease can cause mortalities reaching 100% within 3–10 days of gross signs appearing. During the period of culture, immunostimulant agents and vaccines may provide potential methods to protect shrimps from opportunistic and pathogenic microrganisms. In this study, firstly, WSSV was isolated from infected shrimp and then multiplied in crayfish. WSSV was purified from the infected crayfish haemolymph by sucrose gradient and confirmed by transmission electron microscopy. In vivo virus titration was performed in shrimp, Penaeus semisulcatus. The LD50 of live virus stock was calculated 10 5.4 /mL. Shrimp post-larvae(1–2 g) were treated with gamma-irradiated(different doses) WSSV(10 0 to 10-4 dilutions) for a period of 10 days. The dose/survival curve for irradiated and un-irradiated WSSV was drawn; the optimum dose range for inactivation of WSSV and unaltered antigenicity was obtained 14–15 kGy. This preliminary information suggests that shrimp appear to benefit from treatment with gammairradiated WSSV especially at 14–15 KGy.     

Effects of virus infection on expression of cell cycle regulatory proteins in the unicellular marine algae Emiliania huxleyi

The authors have investigated the biochemical events by which marine algal virus infection induces cell cycle arrest. The key G₂/M-phase regulatory proteins are analyzed by immunobloting in unicellular Emiliania huxleyi, suggesting that virus induced cell cycle arrest is related with virus''s effect on cyclins and cyclin dependent kinases. E. huxleyi virus (EhV) represses Cdc2/cyclinB complex activity by inhibiting the activity of Cdc2 kinase in a phosphorylation-related manner, blocking host cells G₂/M checkpoint. Dephosphorylated/inactive Cdc25C combined with up-regulation of Wee1 expression at early infect period appears to be important mechanisms by which EhV represses Cdc2/cyclinB complex activity that is required for entry into M phase. This study has allowed us to confirm that algal virus infection leads to selective activation or inhibition of certain cell-cycle factors, which may play a significant role in establishing a more efficient environment for viral gene expression and DNA replication.     

猪传染性胃肠炎病毒S、N基因核酸疫苗的构建与免疫原性试验

采用RT—PCR扩增TGEVSCH株S基因5’端主要抗原位点编码区和全长N基因,将其插入真核表达载体pVAXl,构建单独与嵌合表达S、N基因的pVAX—S、pVAX—N、pVAX—S—N3种重组质粒。通过脂质体将重组质粒体外转染COS-7细胞,利用间接免疫荧光检测目的基因的表达情况。将6周龄NIH小鼠随机分为5组,分别于腿部肌肉注射重组质粒pVAX、pVAX—S、pVAXS-N、pVAX—S＋pVAX—N与pVAX—S＋pVAX,共免疫3次,间隔2周。通过间接EI。IsA以及流式细胞仪分别检测免疫小鼠血清中特异性TGEVIgG抗体和外周血T淋巴细胞亚群数量。结果显示,重组质粒构建正确且在COS-7细胞中得以表达;pVAX—S＋pVAX—N质粒混合免疫组与pVAX—s＋pVAx质粒混合免疫对照组相比,可有效提高免疫小鼠外周血T淋巴细胞亚群数量和血清中特异性抗体水平的增加,而pVAX—S-N免疫组与pVAX—S组比较,未能起到提高免疫小鼠血清中特异性抗体水平的和外周血T淋巴细胞亚群数量作用。以上结果表明S、N基因的联合应用可有效加强TGEV核酸疫苗的免疫效力,但这种增强作用与S、N基因联合应用的方式直接相关。     

烟草普通花叶病与气象条件的关系

分析了气象条件对闽西北烟草普通花叶病(TMV)发生发展的影响。分析得出:温度、湿度、降水及日照等气象因子对TMV发生发展均有明显的影响,但影响程度和关键时段不尽相同,影响最大的是湿度和降水,温度次之,日照最小;TMV始发时温度起主要作用,而流行时起主要作用的是湿度和降水。建立了闽西北地区TMV发生程度的中期预测模式,并提出了防治措施。     

Influence of Virus upon the Marine Bacterial Metabolism and Its Biogeochemical Effects

Viruses are by far the most abundant entities in marine environments, and are mainly phages that infect bacteria and archaea, which also are a significant component of marine ecosystem and a major force behind marine biogeochemical cycles. As a major source of mortality, viral lysis can release highly labile cellular components, both organic matters and inorganic nutrients, regulating the metabolism of its hosts and influencing the biogeochemical cycles. During infection, viruses could hijack the metabolic system of hosts for its own propagation, thereby changing the metabolism and metabolites of host cells. This paper summarized the effects of viruses on the metabolism of marine bacterioplankton at both the cellular and community level, and its influence on the cycling of ocean elements. Then, the potential impact of environmental factors was assessed on the influence of viruses upon bacterial metabolism. This paper will contribute to a comprehensive understanding of the role of microbes within marine biogeochemical cycles.     

高位池养殖对虾携带白斑综合症病毒变化

采用实时荧光定量 PCR 技术研究6口高位池塘斑节对虾和凡纳滨对虾携带WSSV变化,结果表明:养殖过程中凡纳滨对虾WSSV携带量最高为9.7×105拷贝/g;斑节对虾最高携带量为9.5×105拷贝/g.凡纳滨对虾WSSV感染率分别为:苗种没有携带WSSV;30d为80.0%;60 d为90.O%;90 d为90.0%;120 d为93.3%,斑节对虾潜伏感染率分别为:苗种没有携带WSSV;30d为73.3%;60d为83.3%;90d为90.O%:120d为96.7%.