黄地老虎颗粒体病毒DNA片段的克隆与颗粒体蛋白基因的定位

用ｐＵＣ１９质粒作载体,克隆了黄地老虎颗粒体病毒（Ａｇｒｏｌｉｓｓｅｇｅｔｕｍｇｒａｎｕｌｏｓｉｓｖｉｒｕｓ,简称ＡｓＧＶ）ＤＮＡＰｓｔＩ－Ｄ．Ｅ．Ｆ．Ｇ．Ｈ．Ｊ．Ｋ．等７个片段。以［￣（３２）Ｐ］－ｄＣＴＰ标记的油桐尺蠖核型多角体病毒（Ｂｕｚｕｒａｓｕｐｐｒｅｓｓａｒｉａｎｕｃｌｃａｒｐｏｌｙｈｅｄｒｏｓｉｓｖｉｒｕｓ简称ＢｓＮＰＶ）多角体蛋白基因为探针,在３７℃条件下对ＡｓＧＶ）颗粒体蛋白基因进行了定位,将其分别定位于ＢｓｌⅡ－Ｓ或ＴＰｓＴＩ－Ａ或Ｂ和ＥｃｉＲＩ－Ａ片段上。     

寄主防卫过程中病毒的维管积集与传播

长期以来 ,对病毒侵染的研究 ,在病毒复制及病毒的细胞间转移方面相对要深入一些[1] ,但对病毒的组装、依赖于维管系统的长距离运输等的认识非常有限[2 ] 。对寄主防卫系统及其机制的认识则主要限于一些抗性因子的证实 ,如抗ＴＭＶ的烟草上分离的Ｎ基因 ,抗ＰＶＡ的马铃薯上分离的Ｒ基因 ,以及一些病程相关蛋白因子 ,如几丁质酶Ａ(ＣｈｔＡ)和ＰＲ 1 0ａ等[2～4 ] 。借助于一些标记蛋白 ,如β ｇｌｕｃｕｒｏｎｉｄａｓｅ(Ｇｕｓ)、Ｌｉｃｉｆｅｒａｓｅ(Ｌｉｃｉ)和ｇｒｅｅｎｆｌｕｏｒｅｓｃｅｎｔｐｒｏｔｅｉｎ(ＧＦＰ)等 ,通过跟踪嵌…     

中国对虾一种球状病毒的分离提纯及其核酸蛋白特性的研究

从感染致病的中国对虾（Ｐｅｎａｅｕｓｃｈｉｎｅｓｉｓ）中分离到一种球状病毒,其育径约为２０±４ｎｍ。进行人工感染实验,对虾死亡率为６６％;用脱氧核糖核酸酶（ＤＮａｓｅ）,核糖核酸酶（ＲＮａｓｅ）及二苯胺染色法对病毒核酸进行处理,证明该病毒核酸为脱氧核糖核酸,用Ｓｌ核酸酶（Ｓｌｎｕｃｌｅａｓｃ）对该核酸进一步消化处理,进一步证实该病毒含单链ＤＮＡ。ＳＤＳ－ＰＡＧＥ结果显示,该病毒含４条结构多肽,其分子量分别为８６ｋＤ,７９ｋＤ,７０ｋＤ及２５．５ｋＤ。根据上述特性分析,该病毒可能属于细小病毒科（ｐａｒ－ｖｏｖｉｒｉｄａｅ）,故暂定名为中国对虾细小病毒（ＰｅｎａｅｕｓｃｈｉｎｅｓｉｓＰａｒｖｏｖｉｒｕｓ简称ＰｃＰＶ）。     

马铃薯卷叶病毒（PLRV）基因组研究进展

马铃薯卷叶病毒（ＰＬＲＶ）基因组研究进展张鹤龄（内蒙古大学生物系,呼和浩特０１００２１）关键词马铃薯卷叶病毒,基因组ＴｈｅＡｄｖａｎｃｅｓｉｎＰＬＲＶＧｅｎｏｍｅＲｅｓｅａｒｃｈＺｈａｎｇＨｅｌｉｎｇ（ＩｎｎｅｒＭｏｎｇｏｌｉａＵｎｉｖｅｒｓｉｔｙ,...     

真核藻类的病毒和病毒类粒子（VLPs）

真核藻类的病毒和病毒类粒子（ＶＬＰｓ）赵以军石正丽（中国科学院水生生物研究所,武汉４３００７２）（中国科学院武汉病毒研究所,武汉４３００７１）ＶｉｒｕｓｅｓａｎｄＶｉｒｕｓ－ｌｉｋｅＰｅａｒｔｉｃｌｅｓｏｆＥｕｋａｒｙｏｔｉｃＡｌｇａｅＺｈａｏＹｉｊ...     

猪瘟病毒石门株NS2—3基因片段的序列测定及比较

根据猪瘟病毒Ｃ株的序列,以计算机辅助设计,化学合成１对引物（ＰＦ５６４８／ＰＲ６６０４）,应用ＲＴＰＣＲ技术从感染猪血中成功地扩增了我国猪瘟病毒强毒石门株ＮＳ２３基因片段,大小为９５７ｂｐ,位于ＮＳ３基因的中部ＮＴＰａｓｅ和Ｈｅｌｉｃａｓｅ活性区。克隆后测序,结果表明该段基因产物具有解旋酶超家族全部七个特征性保守序列,包括共同的ＮＴＰ结合基序Ａ位点（ＧＸＧＫＴ／Ｓ）和Ｂ位点（３ｈｙ,２ｘ）Ｄ。序列同源性比较表明,石门株与日本的ＡＬＤ和ＧＰＥ－株同源性最高,与其它３株猪瘟病毒（Ｃ株、Ｂｒｅｓｃｉａ株和Ａｌｆｏｒｔ株）的同源性也很高,并与２株牛病毒性腹泻病毒（ＢＶＤＶ）（ＮＡＤＬ株和ＳＤ１株）也有较高的同源性,尤其是由核苷酸序列推导的氨基酸序列,同源性均大于９０％,是瘟病毒属基因组中最保守的区段,这与该基因产物在病毒复制及聚蛋白前体加工过程中所具有的重要功能是一致的     

植物病毒卫星研究进展

植物病毒卫星研究进展周雪平,李德葆（浙江农业大学生物技术研究所,杭州３１００２９）自６０年代Ｋａｓｓａｎｉｓ发现有些烟草坏死病毒（ＴＮＶ）分离物中含有１７ｎｍ的病毒颗粒,并将其称谓卫星烟草坏死病毒（ＳＴＮＶ）以来,已发现多种植物ＲＮＡ病毒含有卫星（Ｓ...     

杆状病毒增效蛋白研究进展

昆虫杆状病毒增效蛋白（ｅｎｈａｎｃｉｎ）曾被称为病毒促进因子（ｓｙｎｅｒｇｉｓｔｉｃｆａｃｔｏｒ,ＳｙＦ）或病毒增效因子（ｖｉｒａｌｅｎｈａｎｃｉｎｇｆａｃｔｏｒ,ＶＥＦ）,它长期以来被认为只存在于昆虫颗粒体病毒（Ｇｒａｎｕｌｏｓｉｓｖｉｒｕｓ,ＧＶ）中的一类蛋白质,已知有８种ＧＶ中含有增效蛋白。自Ｔａｎａｄａ［１～４］１９５９年从美洲一星粘虫（Ｐｓｅｕｄａｌｅｔｉａｕｎｉｐｕｎｃｔａ,Ｐｕ）ＧＶ中分离出增效蛋白并证实它有增强ＰｕＭＮＰＶ的感染力后,一直很受人们关注。从它的作用机理到氨基酸序列…     

两例丙型肝炎病毒(HCV)和乙型肝炎病毒(HBV)重叠感染者的HCV核心区cDNA序列分析

从临床肝病患者中选择两例ＨＣＶ和ＨＢＶ重叠感染者ＨＳＱ和ＳＺＨ,他们血清中的生化指标丙氨酸转氨酶（ＡＬＴ）持续异常,肝活检病理示有严重的肝损伤。在ＡＬＴ异常期,血清学检测结果为ＨＢｓＡｇ、ＨＢｅＡｇ阳性,抗ＨＣＶＩｇＧ（包括Ｃ２２、Ｃ３３ｃ）阴性,但套式ＰＣＲ检测ＨＣＶＲＮＡ阳性,核心区ｃＤＮＡ序列分析发现该区有１个密码子（ＧＧＣｎｔ３８５—３８７）缺失,对应缺失的氨基酸是甘氨酸（ＧＬＹ）,从血清学检测和序列分析结果推测,在ＨＣＶ和ＨＢＶ重叠感染中,ＨＢＶ和ＨＣＶ均可处于持续复制状态,抗ＨＣＶＩｇＧ抗体阴性可能是ＨＣＶ的多蛋白前体翻译和病毒颗粒装配受到ＨＢＶ干扰的结果。     

兔出血症病毒信使RNA的分离纯化及其生物活性的鉴定

用液相免疫沉法从感染兔出血症病毒（Ｒａｂｂｉｔ Ｈａｅｍｏｒｒｈａｇｉｃ Ｄｉｓｅａｓｅ Ｖｉｒｕｓ,ＲＨＤＶ）８小时后的兔肝组织的多聚核糖体中,分离出ＲＨＤＶ特异性的多聚核糖体,并由此获得全核酸,经Ｏｌｉｇｏ（ｄＴ）－ｃｅｌｌｕｌｏｓｅ亲和层板,得到３．４Ａ２６０的ＲＨＤＶｐｏｌｙ（Ａ）＾＋ＲＮＡ占全核酸的３．５％。此Ｐｏｌｙ（Ａ）＾＋ＲＮＡ经甲醛变性琼脂糖电泳,得到６条单一的带,其大小分别为０     

A new nodavirus-negative Trichoplusia ni cell line for baculovirus-mediated protein production

Cell lines derived from Trichoplusia ni (Tn) are widely used as hosts in the baculovirus-insect cell system (BICS). One advantage of Tn cell lines is they can produce recombinant proteins at higher levels than cell lines derived from other insects. However, Tn cell lines are persistently infected with an alphanodavirus, Tn5 cell-line virus (TnCLV), which reduces their utility as a host for the BICS. Several groups have isolated TnCLV-negative Tn cell lines, but none were thoroughly characterized and shown to be free of other adventitious viruses. Thus, we isolated and extensively characterized a new TnCLV-negative line, Tn-nodavirus-negative (Tn-NVN). Tn-NVN cells have no detectable TnCLV, no other previously identified viral contaminants of lepidopteran insect cell lines, and no sequences associated with any replicating virus or other viral adventitious agents. Tn-NVN cells tested negative for >60 species of Mycoplasma, Acholeplasma, Spiroplasma, and Ureaplasma. Finally, Tn-NVN cells grow well as a single-cell suspension culture in serum-free medium, produce recombinant proteins at levels similar to High Five™ cells, and do not produce recombinant glycoproteins with immunogenic core α1,3-fucosylation. Thus, Tn-NVN is a new, well-characterized TnCLV-negative cell line with several other features enhancing its utility as a host for the BICS.     

Counter Current Line Absorption Immunoelectrophoresis is an Alternative Diagnostic Screening Test to Counter Current Immunoelectrophoresis in Aleutian Disease (AD) Eradication Programs

Counter current immunoelectrophoresis (CCIE) is the diagnostic method used in the ongoing Aleutian disease virus eradication program on Danish mink farms. There has been an increasing demand for an alternative diagnostic test especially to evaluate suspected false positive CCIE reactions. We compared test results of a number of negative and positive mink sera in indirect counter current immunoelectrophoresis (ICCIE), counter current line absorption immunoelectrophoresis (CCLAIE) and radio immuno assay (RIA) with test results from counter current immunoelectrophoresis and found that counter current line absorption immunoelectrophoresis is the best alternative diagnostic screening test to counter current immunoelectrophoresis for Aleutian disease eradication programs. Not only proved the CCLAIE test to be useful for evaluation of doubtfully positive CCIE reactions, but it was found to have a higher sensitivity than the CCIE test.     

我国南方花生发生一种由蕃茄斑萎病毒引起的新病害

我们于1984和1985年6月上、中旬,在广州市郊、县,湛江市郊以及广西南宁市郊、县,北海市郊和合蒲县等花生产区,调查花生病毒病时,除了发现花生轻斑驳病毒病外,还发现一种新的病毒病害。其症状特征是:病株顶端叶片上出现很多褪绿黄斑或环斑,有的环斑变     

Efficient method to isolate and purify viruses of bacteria from marine environments

Aim:  To isolate viruses of specific heterotrophic bacterial strains from marine environments using a host addition/virus amplification protocol (HAVAP) for use in phage/host systems.
Methods and Results:  Bacteria-free seawater samples containing natural viruses assemblages were inoculated with a single laboratory grown bacterial host of interest in a nutrient-enriched [peptone, Fe(III) and yeast extract] seawater suspension. These conditions enhanced the replication of only those virus(s) capable of infecting the host bacterium. After incubation, free viruses were recovered at concentrations ranging 10⁵–10¹⁰ infectious virus particles per ml of seawater. Using this approach, 15 viruses were isolated and represented 12 unique phage/host systems. Two of the hosts tested were infected by more than one virus.
Conclusions:  Isolation of high concentrations of specific viruses is possible even if their initial concentrations in native waters are low. This approach allows the recovery of phage/host systems that may not be numerically dominant.
Significance and Impact of the Study:  This host enrichment protocol for virus detection and isolation is well-suited for aquatic viral ecology studies that require phage/host systems.     

Ribosome inactivating proteins from plants inhibiting viruses

Many plants contain ribosome inactivating proteins (RIPs) with N-glycosidase activity, which depurinate large ribosomal RNA and arrest protein synthesis. RIPs so far tested inhibit replication of mRNA as well as DNA viruses and these proteins, isolated from plants, are found to be effective against a broad range of viruses such as human immunodeficiency virus (HIV), hepatitis B virus (HBV) and herpes simplex virus (HSV). Most of the research work related to RIPs has been focused on antiviral activity against HIV; however, the exact mechanism of antiviral activity is still not clear. The mechanism of antiviral activity was thought to follow inactivation of the host cell ribosome, leading to inhibition of viral protein translation and host cell death. Enzymatic activity of RIPs is not limited to depurination of the large rRNA, in addition they can depurinate viral DNA as well as RNA. Recently, Phase I/II clinical trials have demonstrated the potential use of RIPs for treating patients with HIV disease. The aim of this review is to focus on various RIPs from plants associated with anti-HIV activity.     

Serological study on the transmission of a granulosis virus of the armyworm,Pseudaletia unipuncta (Lepidoptera: Noctuidae), to other lepidopterous species

The synergistic (Hawaiian) strain of a granulosis virus (GV) from the armyworm, Pseudaletia unipuncta, was transmitted to Spodoptera exigua, Autographa californica, and Trichoplusia ni. The viruses isolated from these hosts were tested by radial double-immunodiffusion (RDD) and immunoelectrophoresis (IE) for their relationship to the original virus. Untreated and heat-treated virus inclusion bodies (capsules) were compared for their antigenic properties but no differences were detected. The antiserum elicited against the whole capsule was more sharply specific for the antigenic determinants than the one elicited against the dissolved capsule proteins. The viruses obtained from S. exigua and T. ni elicited precipitin lines that differed from those of the P. unipuncta GV in their electrophoretic mobility with the one-trough IE method; however, with the two-trough method, the lines fused indicating that the antigens were identical. The major precipitin line indicating identity of the viruses wasthat produced by the synergistic factor (SF) purified from the capsule proteins of the synergistic GV strain. The presence of SF in the GV produced in alternate hosts indicated that its production was virus directed. The SF was not detected in the GVs of Laspeyresia pomonella and Pieris rapae and in the nonsynergistic Oregonian GV of P. unipuncta. A field-collected GV from S. exigua exhibited a different precipitin pattern from that of the synergistic GV, but one of the precipitin lines shared partial relationship to the SF.     

一种以我国痘苗病毒天坛株血凝素基因为选择标记的新型系列表达载体的构建

本文作者从痘苗病毒天坛株Sal I基因文库中确定并分离了痘苗病毒血凝素(HA)基因并组建了以HA为选择标记的系列表达载体,可用于不同结构和不同要求的外源基因在真核细胞中的表达。本系列载体的优点是,在病毒重组过程中可避开传代细胞系,一次性选择出可供疫苗使用的重组病毒。而HA阴性的重组病毒的毒力在家兔皮内试验中,未见明显下降。     

Virus particle packaging in baculovirus and cytoplasmic polyhedrosis virus inclusion bodies

The structure of the inclusion bodies (IBs) of three multiply enveloped nuclear polyhedrosis viruses (MNPVs), one singly enveloped NPV (SNPV), two granulosis viruses (GVs) and one cytoplasmic polyhedrosis virus (CPV) were compared. A method was devised to calculate the numbers of virus particles and nucleocapsids in IBs using data from light microscopy and thin sections. The three MNPVs, from Agrotis segetum (English and Polish virus isolates) and Mamestra brassicae had similar concentrations of virus particles ranging from 17.3 to 19.6 per μm³ of IB. Plusia gamma SNPV had a higher density of 59.6 virus particles per μm³ of IB, which partly compensated for its having smaller IBs (mean volume 0.65 μm³) than the MNPVs (2.60–9.71 μm³). The English A. segetum MNPV isolate had the most nucleocapsids in each virus particle (mean, 4.04) and the largest IBs (mean volume, 9.71 μm³), giving 674 nucleocapsids per IB on average. The GVs, from A. segetum and Pieris brassicae, mainly contained one nucleocapsid per IB. P. gamma CPV IBs had a much higher density of virus particles than the baculoviruses (260 per μm³ compared with 17–60 per μm³). These data are discussed in relation to the biological properties of these viruses, and possible adaptational advantages of alternative IB designs are considered.     

DEAD-box RNA解旋酶家族在天然免疫应答信号通路中的作用

病毒入侵宿主细胞时,宿主细胞启动抑制病毒复制的免疫机制.同样,病毒也会利用多种手段去逃避先天免疫感应机制的监测以及宿主细胞对外来者的降解,同时还会操纵宿主细胞为自身的增殖提供便利.DEAD-box解旋酶家族是一类存在于宿主细胞中的功能蛋白,它们在转录、剪接、mRNA的合成和翻译等多种细胞过程中起着关键作用.该家族成员拥...     

Host range and virulence of five baculoviruses from lepidopterous hosts

The host range and virulence of five insect baculoviruses (two multiply-enveloped nuclear polyhedrosis viruses (MNPVs) from Agrotis segetum and Mamestra brassicae; one singly-enveloped NPV from Plusia gamma and two granulosis viruses (GVs) from A. segetum and Pieris brassicae) were studied for seven lepidopterous pests of temperate agriculture (A. segetum, Agrotis exclamationis, Lacanobia oleracea, M. brassicae, Noctua pronuba, P. gamma and Pieris rapae). None of the viruses killed all species but M. brassicae MNPV failed to infect only P. rapae. The other viruses were restricted to the homologous host, or members of its genus or subfamily. In all examples except A. segetum GV, the median lethal dose for the most susceptible host, was less than 22 virus inclusion bodies and median lethal times for all infections ranged from 5·5 to 16·6 days. The low susceptibility of A. segetum and other noctuids to GV infections is discussed in relation to the structure of inclusion bodies and the nature of the infectious unit in baculoviruses.