Nonoccluded baculovirus- and filamentous virus-like particles in the spotted cucumber beetle,diabrotica undecimpunctata (coleoptera: chrysomelid)

Nonoccluded baculovirus-and filamentous virus-like particles were found in nuclei of hemocytes or midgut cells of field-collected spotted cucumber beetles. Each type of particle was associated with a different type of virogenic stroma containing various viral components similar to those referred to as capsid, nucleocapsid, viroplasm, and viral envelope in other known baculovirus infections. Nucleocapsids of the virus which occured only in hemocytes were rod-shaped particles approximately 230 nm long and 52 nm wide and were enveloped singly by a trilaminar unit membrane. Enveloped and partly enveloped particles appeared to be released from the nucleus to the cytoplasm by budding through the nuclear envelope acquiring additional membranes. The nucleocapsids of the virus which occurred only in nuclei of midgut cells were filamentous particles with an average diameter of 25 nm and variable length up to 2 μm. Some extremely long particles were bent almost 360° near the middle, resulting in a hairpin-like configuration. The particles were always enveloped singly. No particles budding through the nuclear envelope were observed.     

The rabbit C family of short, interspersed repeats. Nucleotide sequence determination and transcriptional analysis

When the entire adeno-associated virus (AAV) genome is inserted into a bacterial plasmid, infectious AAV genomes can be rescued and replicated when the recombinant AAV-plasmid DNA is transfected into human 293 cells together with helper adenovirus particles. We have taken advantage of this experimental system to analyze the effects of several classes of mutations on replication of AAV DNA. We obtained AAV mutants by molecular cloning in bacterial plasmids of naturally occurring AAV variant or defective-interfering genomes. Each of these mutants contains a single internal deletion of AAV coding sequences. Also, some of these mutant-AAV plasmids have additional deletions of one or both AAV terminal palindromes introduced during constructions in vitro. We show here that AAV mutants containing internal deletions were defective for replicative form DNA replication (rep-) but could be complemented by intact wild-type AAV. This indicates that an AAV replication function, Rep, is required for normal AAV replication. Mutants in which both terminal palindromes were deleted (ori-) were also replication defective but were not complementable by wild-type AAV. The cis-dominance of the ori- mutation shows that the replication origin is comprised in part of the terminal palindrome. Deletion of only one terminal palindrome was phenotypically wild-type and allowed rescue and replication of AAV genomes in which the deleted region was regenerated apparently by an intramolecular correction mechanism. One model for this correction mechanism is proposed. An AAV ori- mutant also complemented replication of AAV rep- mutants as efficiently as did wild-type AAV. These studies also revealed an unexpected additional property of the deletion mutants in that monomeric single-stranded single-stranded DNA accumulated very inefficiently even though monomeric single-stranded DNA from the complementing wild-type AAV did accumulate.     

Decreased High-Affinity Binding of [3H]Muscimol to Cerebral Synaptic Membranes of Scrapie-Infected Mice

Scrapie is a transmissible disease that results in progressive degeneration of the central nervous system and death. Although scrapie has been studied histopathologically, relatively little is known concerning neurotransmitter alterations. Specific [3H]muscimol binding to whole brain crude synaptic membranes (CSM) from mice clinically affected with scrapie was significantly (p less than 0.01) reduced, to approximately 73% of that of the controls. Of the brain regions examined, binding to only cerebral CSM was significantly (p less than 0.0001) decreased. Scatchard analyses of saturation curves revealed that the high-affinity (KD = 8 +/- 3 nM) site for muscimol was abolished in cerebral CSM from scrapie-infected mice, while the low-affinity site was unaffected. Binding of [3H]flunitrazepam to cerebral CSM was unaffected by scrapie and was stimulated by GABA to the same extent in both scrapie and control mice. These results suggest that scrapie agent 139A in C57BL/6J mice manifests a portion of its CNS pathology via a high-affinity GABA binding site that is unassociated with the benzodiazepine receptor.     

Structure of tomato bushy stunt virus: V. Coat protein sequence determination and its structural implications

We report the chemically determined sequence of most of the polypeptide chain of the coat protein of tomato bushy stunt virus. Peptide locations have been determined by comparison with the high-resolution electron density map from X-ray crystallographic analysis as well as by conventional chemical overlaps. Three small gaps remain in the 387-residue sequence. Positively charged side-chains are concentrated in the N-terminal part of the polypeptide (the R domain) as well as on inward-facing surfaces of the S domain. There is homology of S-domain sequences with structurally corresponding residues in southern bean mosaic virus.     

Complete nucleotide sequences of all three poliovirus serotype genomes. Implication for genetic relationship, gene function and antigenic determinants

The complete nucleotide sequences of the genomes of the type 2 ( P712 , Ch, 2ab ) and type 3 (Leon 12a1b ) poliovirus vaccine strains were determined. Comparison of the sequences with the previously established genome sequence of type 1 (LS-c, 2ab ) poliovirus vaccine strain revealed that 71% of the nucleotides in the genome RNAs were common, that the 5' and 3' termini of the genomes were highly homologous, and that more than 80% of the nucleotide differences in the coding region occurred in the third letter position of in-phase codons, resulting in a low frequency of amino acid difference. These results strongly suggested that the serotypes of poliovirus derived from a common prototype. A comparison of the amino acid sequences predicted from the genome sequences showed highest variation in the capsid protein region, whereas non-structural proteins are highly conserved. Initiation of polyprotein synthesis occurs in all three strains more than 740 nucleotides downstream from the 5' end. An analysis of the non-coding region suggests that small peptides that could potentially originate from this region are conserved. The amino acid sequences immediately surrounding the cleavage signals, however, show a higher than average degree of variation. The analysis of the amino acid sequences of the capsid protein VP1 of all serotypes has led to the prediction of potential antigenic sites on the virion involved in neutralization.     

杨尺蠖核型多角体病毒蛋白质的特性

本文研究了杨尺蠖核型多角体病毒(AciNPV)蛋白质特性,用反复调等电点法,从AciNPV的多角体蛋白中分离到A,B两种蛋白,Sepharose 6B柱层析和超离心沉降分析表明,两者均为一个峰纯,沉降系数(S20w)分别为4.9和8.9,对A蛋白进行了16种氨基酸组成分析,Glu、Asp和Leu含量高于其它氨基酸,不含Cys,用SDS-聚丙烯酰胺凝胶电泳和免疫双扩散法分析表明,用两倍体积饱和硫酸铵沉淀法制备的多角体蛋白,与提纯的多角体A、B蛋白之间无差异,多角体蛋白结构多肽由6种组成,分子量在12500—54000道尔顿范围内,其中32000是多角体蛋白的主要多肽,病毒粒子结构多肽和核衣壳蛋白分别由19和7个多肽组成,分子量范围分别在89000～13000和34000～13000之间,间接试验结果表明,在AciNPV中有碱性蛋白酶活性存在。     

美味侧耳病毒及其生化特性

自美味侧耳(Pleurotus sapidus)子实体中分离到两类病毒颗粒,一是球形,直径为25nm(少数为19、32am);一是杆状,具较深锯齿状亚基,大小为12～14×40～600nm,病毒具核蛋白吸收峰,最大吸收为E257nm,最小为242nm,ISCO蔗糖密度梯度离心中出现3个峰。 病毒具有分子量为44000和22000两条主要衣壳多肽,自子实体中提取到分子量约1.2×10~(?)道尔顿的dsRNA,可能是球形病毒的基因组,病毒与同属真菌糙皮侧耳(P,ostreatus)和德平菇(P,floride)球形病毒之间有血清学关系。     

流行性乙型脑炎病毒血清抗体半微量空斑抑制法的改进

本文报道一种检测流行性乙型脑炎(简称乙脑)中和抗体的简化空斑抑制试验方法,用BHK21传代细胞在24孔塑料板上进行试验,在细胞未形成单层时,将病毒直接接种在刚消化的细胞悬液内,孵育后加甲基纤维素复盖物,再培养,细胞层经染色后即可计算空斑数。 用该法检查47名儿童乙脑疫苗免疫前后的中和抗体,与常规的鸡胚细胞全量法或BHK21传代细胞微量中和试验法作了比较,结果表明本法与其它常规法的敏感性一致,且具有简便快速、节省人力和原材料、空斑形成率高的优点,便于大量血清学检测。     

大尺蠖核多角体病毒DNA的限制性消化和物理图谱

用5种限制性核酸内切酶消化大尺蠖核多角体病毒(BsNPV)基因组DNA,所得片段数分别是:BamH Ⅰ,9;Bgl Ⅱ,7;Xho Ⅰ,10;HindⅢ 13;EcoR Ⅰ,12,从各个片段的累加测出BsNPV基因组的平均大小约为91,75kb;分子量约为59.60×10~6d。在单酶消化的基础上,用BamH Ⅰ/Bgl Ⅱ双酶消化得到16个片段(即9+7);大小为91,27kb,用、Bgl Ⅱ/Xho Ⅰ双酶消化产生7+10=17个片段,大小为90.04kb,由此组建了这三个酶在BsNPV基因组上的物理图谱。     

流行性成人腹泻病人排出成人腹泻轮状病毒的持续时间

成人腹泻轮状病毒(Adult Diarrhoea Rotavirus,简称ADRV)是引起青壮年急性胃肠炎的主要病原之一,1983年至1985年在我国许多地区暴发流行,严重危害着人民健康,至今尚无治疗药物。因此,摸清病人排病毒情况或规律,对及时控制传染源,预防ADRV大量传播有着重要意义。现将本文实验结果报告如下。