伪狂犬病毒闽A株基因文库的构建及物理图谱分析

本文报道以质粒ｐＢＲ３２２作载体,用鸟枪法克隆出了ＰＲＶ闽Ａ株除ＢａｍＨＩ－１,２外的所有酶切片段,构建了ＰＲＶ闽Ａ株基因文库,并以克隆出的ＢａｍＨＩ片段用光生物素标记作探针,应用分子杂交法确定了ＰＲＶ闽Ａ株绝大部分限制性内切酶位点的位置。     

HSV-1UL6基因编码门蛋白的结构及功能

门蛋白在单纯疱疹病毒1型（HSV-1）的衣壳装配和基因组包装过程中扮演重要角色,对病毒的复制有重要作用。本文就HSV-1 UL6基因编码的门蛋白进行阐述,具体介绍形成门蛋白多聚体的影响因素,门蛋白及其周边蛋白的结构及功能,以及其靶点药物的研究现状等,以期为研究门蛋白在疱疹病毒感染和复制过程中的作用提供一定参考。     

Characterization of subcellular localization of duck enteritis virus UL51 protein

Background

Dengue virus (DENV), a mosquito borne flavivirus is an important pathogen causing more than 50 million infections every year around the world. Dengue diagnosis depends on serology, which is not useful in the early phase of the disease and virus isolation, which is laborious and time consuming. There is need for a rapid, sensitive and high throughput method for detection of DENV in the early stages of the disease. Several real-time PCR assays have been described for dengue viruses, but there is scope for improvement. The new generation TaqMan Minor Groove Binding (MGB) probe approach was used to develop an improved real time RT-PCR (qRT-PCR) for DENV in this study.

Results

The 3'UTR of thirteen Indian strains of DENV was sequenced and aligned with 41 representative sequences from GenBank. A region conserved in all four serotypes was used to target primers and probes for the qRT-PCR. A single MGB probe and a single primer pair for all the four serotypes of DENV were designed. The sensitivity of the two step qRT-PCR assay was10 copies of RNA molecules per reaction. The specificity and sensitivity of the assay was 100% when tested with a panel of 39 known positive and negative samples. Viral RNA could be detected and quantitated in infected mouse brain, cell cultures, mosquitoes and clinical samples. Viral RNA could be detected in patients even after seroconversion till 10 days post onset of infection. There was no signal with Japanese Encephalitis (JE), West Nile (WN), Chikungunya (CHK) viruses or with Leptospira, Plasmodium vivax, Plasmodium falciparum and Rickettsia positive clinical samples.

Conclusion

We have developed a highly sensitive and specific qRT-PCR for detection and quantitation of dengue viruses. The assay will be a useful tool for differential diagnosis of dengue fever in a situation where a number of other clinically indistinguishable infectious diseases like malaria, Chikungunya, rickettsia and leptospira occur. The ability of the assay to detect DENV-2 in inoculated mosquitoes makes it a potential tool for detecting DENV in field-caught mosquitoes.     

A Genome Scan for Quantitative Trait Loci Associated with Vibrio anguillarum Infection Resistance in Japanese Flounder (Paralichthys olivaceus) by Bulked Segregant Analysis

A recent genetic linkage map was employed to detect quantitative trait loci (QTLs) associated with Vibrio anguillarum resistance in Japanese flounder. An F1 family established and challenged with V. anguillarum in 2009 was used for QTL mapping. Of the 221 simple sequence repeat (SSR) markers used to detect polymorphisms in the parents of F1, 170 were confirmed to be polymorphic. The average distance between the markers was 10.6 cM. Equal amounts of genomic DNA from 15 fry that died early and from 15 survivors were pooled separately to constitute susceptible bulk and resistance bulk DNA. Bulked segregant analysis and QTL mapping were combined to detect candidate SSR markers and regions associated with the disease. A genome scan identified four polymorphic SSR markers, two of which were significantly different between susceptible and resistance bulk (P?=?0.008). These two markers were located in linkage group (LG) 7; therefore, all the SSR markers in LG7 were genotyped in all the challenged fry by single marker analysis. Using two different models, 11–17 SSR markers were detected with different levels of significance. To confirm the associations of these markers with the disease, composite interval mapping was employed to genotype all the challenged individuals. One and three QTLs, which explained more than 60 % of the phenotypic variance, were detected by the two models. Two of the QTLs were located at 48.6 cM. The common QTL may therefore be a major candidate region for disease resistance against V. anguillarum infection.     

温度升高对高光强环境下蛋白核小球藻（Chlolorella pyrenoidosa）光能利用和生长的阻抑效应

以蛋白核小球藻（Cholorella pyrenoidosa）为实验材料,研究了温度变化对不同光照水平下蛋白核小球藻的光能利用和生长的影响,以明确光照强度对微藻的光能利用和生长的影响是否因温度不同而发生变化。实验中共设置了3个光照强度水平（50,150,300μmol•m^-2•s^-1）和2个温度水平（15℃,25℃）。实验结果表明,不同光照水平下小球藻叶绿素荧光的非光化学淬灭（NPQ）大小与温度有关,光照强度为150,300μmol•m^-2•s^-1时,温度升高使小球藻叶绿素荧光NPQ提高,并且光照强度越高小球藻叶绿素荧光NPQ增大越多,50μmol•m^-2•s^-1光照强度下温度升高对叶绿素荧光NPQ没有影响。实验发现,25℃培养温度下小球藻的光合电子传递速率(ETR)随光照强度增高而上升的速率要低于15℃时小球藻ETR上升的速率;随着光照强度增高,温度升高使小球藻ETR降低程度增大。实验结果还表明,15℃时小球藻培养液叶绿素a浓度随光照强度升高而增高,300μmol•m^-2•s^-1培养光强下具有最高的叶绿素a浓度。但在25℃时,光照强度升高叶绿素a浓度并不一定增高,300μmol•m^-2•s^-1光照强度下的叶绿素a浓度比150μmol•m^-2•s^-1光照强度下要低。本研究表明,温度升高增大了高光照水平下蛋白核小球藻对光能的热耗散,使光照增强对小球藻生长的促进作用减弱。由于温度升高对小球藻光能利用和生长的阻抑作用,小球藻生长的适宜光照水平因温度升高而降低。      

长白山阔叶红松林生长季热量平衡变化特征

根据长白山阔叶红松林2001年5月下旬至10月上旬微气象梯度观测资料和辐射、土壤热通量资料,用波文比-能量平衡方法(BREB方法)计算了森林的显热通量和感热通量,并计算了森林大气和植被体的储热量,分析了阔叶红松林热量平衡各项的日变化和季节变化,结果发现,热量平衡(净辐射)与太阳总辐射呈线性关系;热量平衡各项都与净辐射有相同的日变化特征,为昼正夜负的曲线．各项的绝对值一般表现为净辐射>潜热通量>感热通量>储热变化．受日照时间的影响,6～10月各分量正值的日持续时间逐渐缩短．月平均结果,白天净辐射6月份最大,10月上旬最小,变化于0～527W·m^-2,夜间的净辐射在0～-121W·m^-2．潜热通量白天和夜间分别在0～441、0～-81W·m^-2,感热通量昼夜分别在0～80、0～-26W·m^-2．储热变化则为0～44、0～-26W·m^-2．白天潜热通量占净辐射的比例8～10月逐渐下降,而感热通量和储热变化的比例9～10月明显上升,特别在严霜后2～3d,出现潜热通量比例突减、感热通量比例突增的现象．文中还对通量观测仪器、方法进行了简要分析．