鲤春血症病毒中国分离株糖蛋白基因和氨基酸序列的初步解析

2002～2004年间从国内养殖鲤科鱼类和观赏鱼类中分离出8株鲤春血症病毒(SVCV).根据SVCV参考株全序列,设计引物,用逆转录聚合酶链式反应扩增出8株SVCV糖蛋白编码基因片段,并对扩增产物进行了克隆和序列测定.用生物信息学方法对测得的序列进行分析,结果8个国内分离株的糖蛋白基因序列与参考株的基因序列相似性均在92%以上,8个国内分离株之间基因序列相似性均在97.7%以上;8个国内分离株之间糖蛋白推导出的氨基酸序列相似性均在94.5%以上,与参考株氨基酸序列相似性在92.9%-94.9%之间.系统发育树分析结果表明,SVCV国内分离株与USA株、980451株、980528株和970469株的进化方向一致,与其它SVCV毒株在进化方向上不同.8个毒株有19个共同的酶切位点,推导出的氨基酸序列中有10个亲水区、10个可能的抗原位点和10个跨膜蛋白区域,其峰值基本一致.对SVCV国内分离株的糖蛋白6个功能位点(天冬酰胺糖基化位点、精氨酸-甘氨酸-天冬氨酸序列、酪蛋白激酶Ⅱ磷酸化位点、蛋白激酶C磷酸化位点、酪氨酸磷酸化位点和肉豆蔻酰基化位点)进行了初步分析.     

鲤春血症病毒中国分离株糖蛋白基因和氨基酸序列的初步解析

2002～2004年间从国内养殖鲤科鱼类和观赏鱼类中分离出8株鲤春血症病毒(SVCV)。根据SVCV参考株 全序列,设计引物,用逆转录聚合酶链式反应扩增出8株SVCV糖蛋白编码基因片段,并对扩增产物进行了克隆和 序列测定。用生物信息学方法对测得的序列进行分析,结果8个国内分离株的糖蛋白基因序列与参考株的基因序 列相似性均在92％以上,8个国内分离株之间基因序列相似性均在97．7％以上;8个国内分离株之间糖蛋白推导 出的氨基酸序列相似性均在94．5％以上,与参考株氨基酸序列相似性在92．9％-94．9％之间。系统发育树分析结 果表明,SVCV国内分离株与USA株、980451株、980528株和970469株的进化方向一致,与其它SVCV毒株在进 化方向上不同。8个毒株有19个共同的酶切位点,推导出的氨基酸序列中有10个亲水区、10个可能的抗原位点 和10个跨膜蛋白区域,其峰值基本一致。对SVCV国内分离株的糖蛋白6个功能位点(天冬酰胺糖基化位点、精 氨酸-甘氨酸-天冬氨酸序列、酪蛋白激酶Ⅱ磷酸化位点、蛋白激酶C磷酸化位点、酪氨酸磷酸化位点和肉豆蔻酰 基化位点)进行了初步分析。     

Venezuelan Equine Encephalitis in Panama: Fatal Endemic Disease and Genetic Diversity of Etiologic Viral Strains

Venezuelan equine encephalitis (VEE) is a reemerging, mosquito-borne viral disease of the neotropics that is severely debilitating and sometimes fatal to humans. Periodic epidemics mediated by equine amplification have been recognized since the 1920s, but interepidemic disease is rarely recognized. We report here clinical findings and genetic characterization of 42 cases of endemic VEE detected in Panama from 1961–2004. Recent clusters of cases occurred in Darien (eastern Panama) and Panama provinces (central Panama) near rainforest and swamp habitats. Patients ranged from 10 months to 48 years of age, and the more severe cases with neurological complications, including one fatal infection, were observed in children. The VEE virus strains isolated from these cases all belonged to an enzootic, subtype ID lineage known to circulate among sylvatic vectors and rodent reservoir hosts in Panama and Peru. These findings underscore endemic VEE as an important but usually neglected arboviral disease of Latin America.     

传染性法氏囊病毒的抗原及分子特征

用鸡胚成纤维细胞对来自野外的 5 个传染性法氏囊病毒株 (IBDV-JD1 、 JD2 、 NB 、 HZ1 、 HZ2) 进行分离,测定理化特性、致病性,同时进行血清亚型测定及 A 片段基因组的克隆分析 . 试验所用 5 个法氏囊组织悬液在鸡胚成纤维细胞盲传 2~14 代后适应细胞并产生细胞病变 . 细胞适应的 IBDV 毒株的理化和形态特征与经典传染性法氏囊病毒株一致 . 除 IBDV-HZ1 、 HZ2 属经典 IBDV 血清型外, IBDV-JD1 、 JD2 和 NB 毒株分属不同的血清亚型 . 人工感染实验结果显示,分离的 IBDV 毒株产生与野外病例相似的临床症状和病变,出现法氏囊滤泡髓质的淋巴细胞变性、坏死和消失 . 基因组序列分析显示, IBDV-NB 毒株 A 片段由 3 264 个核苷酸组成,编码由 145 个氨基酸残基组成的 VP5 和由 1 012 个氨基酸残基组成的多聚蛋白 . 与来自 GenBank 的 IBDV Ⅰ型毒株比较, NB 毒株 A 片段编码的多聚蛋白与 JD1 毒株的同源性最高,达 99.5% , VP2 与 JD1 、 CEF94 、 D78 的同源性为 99.8% , VP3 与 JD1 的同源性为 99.2% , VP4 与 JD1 的同源性为 100% , VP5 与 JD1 , HZ2 , P2 , CEF94 , CT , Cu-1 和 D78 毒株的同源性为 99.3%. NB 毒株 VP2 蛋白的第 253 、 280 、 284 位氨基酸残基与 IBDV 变异毒株和经典毒株一致,但不同于 IBDV 超强毒株 . 这些结果暗示 IBDV 的抗原表位是构象依赖性表位, IBDV 血清亚型的形成与 IBDV 弱毒疫苗病毒株密切相关 .     

EvoSNP-DB: A database of genetic diversity in East Asian populations

Genome-wide association studies (GWAS) have become popular as an approach for the identification of large numbers of phenotype-associated variants. However, differences in genetic architecture and environmental factors mean that the effect of variants can vary across populations. Understanding population genetic diversity is valuable for the investigation of possible population specific and independent effects of variants. EvoSNP-DB aims to provide information regarding genetic diversity among East Asian populations, including Chinese, Japanese, and Korean. Non-redundant SNPs (1.6 million) were genotyped in 54 Korean trios (162 samples) and were compared with 4 million SNPs from HapMap phase II populations. EvoSNP-DB provides two user interfaces for data query and visualization, and integrates scores of genetic diversity (Fst and VarLD) at the level of SNPs, genes, and chromosome regions. EvoSNP-DB is a web-based application that allows users to navigate and visualize measurements of population genetic differences in an interactive manner, and is available online at [http://biomi.cdc.go.kr/EvoSNP/]. [BMB Reports 2013; 46(8): 416-421]     

Molecular analyses of the genus Drunella (Ephemeroptera: Ephemerellidae) in the East Asian region

Limnology - The DNA-based identification of aquatic insects, which show high species diversity, is becoming increasingly popular because these insects are excellent indicators of the condition of...     

稻水象甲在浙江和福建省扩散的空间格局分析

应用地统计学方法和GIS技术分析了稻水象甲在浙江和福建省早稻上的空间分布和动态（1993-2001）。结果表明：稻水象甲主要分布于沿海一带,其空间分布格局在所有年份均表现为聚集分布,并且这种分布格局随时间发生变化。在1993年该虫入侵面积大约为392100公顷,在以后的几年中其面积随时间而持续增加,至2001年达最大,为2533900公顷。稻水象甲起初沿海岸线向南扩散,接着向北,现在该虫已入侵约26个县和190个乡镇。通常,该虫在南部的为害重于北部,玉环、乐清、温岭等地发生最重。稻水象甲主要沿公路和水路扩散,这表明该虫的传播为人类活动所致,因此其扩散速率可以通过对国家和地区间的交通货运进行严格的检疫而得到控制。     

口蹄疫病毒株China／99L区段核苷酸序列测定及比较分析

以口蹄疫病毒株China/ 99RNA为模板 ,反转录并扩增目的cDNA ,然后与 pGEM TEasy载体连接并转化JM10 9菌株 ,提取的重组质粒用电泳、PCRampos和EcoR1酶切法鉴定。该毒株与A10、O1K、O1Campos和TW 45毒株的核苷酸序列差异率分别为 15 .2 7%、15 .5 6 %、15 .5 6 %和 15 .49% ;氨基酸序列差异率分别为 8.2 9%、8.76 %、9.2 2 %和 10 .14%。五个毒株的L/P1连接处均为苷氨酸 (Gly) /异亮氨酸 (Ile)。序列比较表明 ,T C、A G和A C转换率较高 ,是点突变的热点核苷酸 ,是影响氨基酸稳定的因素之一。第 43 5 3、95 10 5、10 8 111、146 15 3、16 1 173、183 188和 182 187区域极有可能是L蛋白酶的活性中心 ,第 48位的H、5 1的C、6 5位的E、95位的H、10 9位的H、138位的H、148位的H和 16 5位的E可能是L蛋白酶的活性位点 ,它们在维持蛋白质的空间构像和功能方面具有重要作用     

口蹄疫病毒株China/99L区段核苷酸序列测定及比较分析

以口蹄疫病毒株China/99 RNA为模板,反转录并扩增目的cDNA,然后与pGEM-T Easy载体连接并转化JM109菌株,提取的重组质粒用电泳、PCR ampos和EcoR1酶切法鉴定.该毒株与A10、O\-1K、O1C ampos和TW45毒株的核苷酸序列差异率分别为15.27%、15.56%、15.56%和15.49%;氨基酸序列差异率分别为8.29%、8.76%、9.22%和10.14%.五个毒株的L/P1连接处均为苷氨酸(Gly)/异亮氨酸(Ile).序列比较表明,T-C、A-G和A-C转换率较高,是点突变的热点核苷酸,是影响氨基酸稳定的因素之一.第43-53、95-105、108-111、146-153、161-173、183-188和182-187区域极有可能是L蛋白酶的活性中心,第48位的H、51的C、65位的E、95位的H、109位的H、138位的H、148位的H和165位的E可能是L蛋白酶的活性位点,它们在维持蛋白质的空间构像和功能方面具有重要作用.     

Complete Genome Sequence Analysis of a Natural Reassortant Infectious Bursal Disease Virus in China

A novel isolate of infectious bursal disease virus (IBDV) was designated GX-NN-L. The GX-NN-L IBDV was a very virulent infectious bursal disease virus (vvIBDV) isolated from broiler flocks in Guangxi province, China, in 2011. The GX-NN-L IBDV caused high mortality, immunosuppression, low weight gain, and bursal atrophy in commercial broilers. Here, we report the complete genome sequence of the GX-NN-L IBDV, a reassortment strain with segments A and B derived from very virulent strains and attenuated IBDV, respectively. These findings from this study provide additional insights into the genetic exchange between attenuated and very virulent strains of IBDV and continuous monitoring of the spread of the virus in chicken.     

Molecular epidemiology of Rabbit Haemorrhagic Disease Virus in Australia: when one became many

Rabbit Haemorrhagic Disease Virus (RHDV) was introduced into Australia in 1995 as a biological control agent against the wild European rabbit (Oryctolagus cuniculus). We evaluated its evolution over a 16‐year period (1995–2011) by examining 50 isolates collected throughout Australia, as well as the original inoculum strains. Phylogenetic analysis of capsid protein VP60 sequences of the Australian isolates, compared with those sampled globally, revealed that they form a monophyletic group with the inoculum strains (CAPM V‐351 and RHDV351INOC). Strikingly, despite more than 3000 rereleases of RHDV351INOC since 1995, only a single viral lineage has sustained its transmission in the long‐term, indicative of a major competitive advantage. In addition, we find evidence for widespread viral gene flow, in which multiple lineages entered individual geographic locations, resulting in a marked turnover of viral lineages with time, as well as a continual increase in viral genetic diversity. The rate of RHDV evolution recorded in Australia ?4.0 (3.3–4.7) × 10^?3 nucleotide substitutions per site per year – was higher than previously observed in RHDV, and evidence for adaptive evolution was obtained at two VP60 residues. Finally, more intensive study of a single rabbit population (Turretfield) in South Australia provided no evidence for viral persistence between outbreaks, with genetic diversity instead generated by continual strain importation.     

我国柑桔主要病毒类病害及其无毒化技术研究进展(综述)

本文简述柑桔黄龙病、衰退病、裂皮病及碎叶病病原性质及对生产的危害,概述柑桔无毒化技术的研究进展。     

The Global Spread of Hepatitis C Virus 1a and 1b: A Phylodynamic and Phylogeographic Analysis

Background

Hepatitis C virus (HCV) is estimated to affect 130–180 million people worldwide. Although its origin is unknown, patterns of viral diversity suggest that HCV genotype 1 probably originated from West Africa. Previous attempts to estimate the spatiotemporal parameters of the virus, both globally and regionally, have suggested that epidemic HCV transmission began in 1900 and grew steadily until the late 1980s. However, epidemiological data suggest that the expansion of HCV may have occurred after the Second World War. The aim of our study was to elucidate the timescale and route of the global spread of HCV.

Methods and Findings

We show that the rarely sequenced HCV region (E2P7NS2) is more informative for molecular epidemiology studies than the more commonly used NS5B region. We applied phylodynamic methods to a substantial set of new E2P7NS2 and NS5B sequences, together with all available global HCV sequences with information in both of these genomic regions, in order to estimate the timescale and nature of the global expansion of the most prevalent HCV subtypes, 1a and 1b. We showed that transmission of subtypes 1a and 1b “exploded” between 1940 and 1980, with the spread of 1b preceding that of 1a by at least 16 y (95% confidence interval 15–17). Phylogeographic analysis of all available NS5B sequences suggests that HCV subtypes 1a and 1b disseminated from the developed world to the developing countries.

Conclusions

The evolutionary rate of HCV appears faster than previously suggested. The global spread of HCV coincided with the widespread use of transfused blood and blood products and with the expansion of intravenous drug use but slowed prior to the wide implementation of anti-HCV screening. Differences in the transmission routes associated with subtypes 1a and 1b provide an explanation of the relatively earlier expansion of 1b. Our data show that the most plausible route of the HCV dispersal was from developed countries to the developing world. Please see later in the article for the Editors'' Summary     

BVD Virus Antigens in Tissues of Persistently Viraemic,Clinically Normal Cattle: Implications for the Pathogenesis of Clinically Fatal Disease

The cellular events involved in precipitation of the clinically fatal outcome of an infection with bovine viral diarrhoea virus (BVDV) remain unresolved, though it is now known that this course of the infection, Mucosal Disease (MD), only occurs in calves persistently infected with non-cytopathic BVDV. In studies aimed at elucidating the pathogenesis of MD, the distribution of BVDV antigens and infectious virus in tissues of persistently infected, clinically normal calves was investigated. Virus antigen was detected in most tissues, in epithelial and immune cells. No signs of an inflammatory response were detected and cytopathological changes were subtle or absent. The infection may nevertheless create a cell-environment which will enhance replication of cytopathic virus. Variations in the clinical, pathomorphologies and virological appearance of MD-cases may depend on both the host-reactions, including virus-induced immunopathology, and the virus-strain combinations in a putative mixed infection.