Molecular characterization of the complete genome of a street rabies virus WH11 isolated from donkey in China

The complete genomic sequence of a rabies virus isolate WH11, isolated from brain tissue of a rabid donkey in China, was determined and compared with other rabies viruses. This is the first Chinese street strain which was isolated from donkey and the entire length and organization of the virus was similar to that of other rabies viruses. Multiple alignments of amino acid sequences of the nucleoprotein, phosphoprotein, matrix protein, glycoprotein, and large protein of WH11 with those of other rabies viruses were undertaken to examine the conservative degree of functional regions. Phylogenetic analysis using the complete genomic sequence of WH11 determined that this isolate is most closely related with rabies viruses previously isolated in China and the attenuated Chinese vaccine strain CTN181.     

Complete genome sequences of three rabies viruses isolated from rabid raccoon dogs and a cow in Korea

The complete genomes of three rabies viruses (BD0406CC, BV9901PJ, and 08F40) of two raccoon dogs (Nyctereutes procyonoides koreensis) and a cow were determined. The genomic organization is typical of rabies viruses, and the open reading frames of the N, P, M, G, and L genes are 1,353, 894, 609, 1,575, and 6,384 bases in length, respectively. The full genome length of the three strains was 11,928 nucleotides, and the sequence similarity between the rabies viruses at the nucleotide level was 98.5–99.5 %. Sequence comparisons indicated that these rabies viruses belong to the “Arctic and Arctic-like” group, with high homology to the Eurasian cluster. All Korean strains were clustered with the Mongolia strains, which belong to Arctic-like 1 clade. The 08F40 and BD0406CC strains were constructed with rabies virus strains isolated in Gangwon province. The BV9901PJ strain was closely related to strains isolated in Gyeonggi province in Korea. Three strains were more dependent upon geographical distribution and time period than host species. Complete genome sequencing of different host-origin rabies viruses will provide information that should contribute to understanding the transmission cycle and genetic variability of rabies from different hosts.     

Comparison of complete amino acid sequences and receptor-binding properties among 13 serotypes of hemagglutinins of influenza A viruses

We determined the sequences of 7 serotypes (H4, H6, H8, H9, H11, H12, and H13) of hemagglutinin (HA) genes, which have not been reported so far. The coding regions consisted of 1692 nucleotides in H4, 1698 in H6, 1695 in H8, 1680 in H9, 1695 in H11, 1692 in H12, and 1698 in H13, and specified 564, 566, 565, 560, 565, 564, and 566 amino acids, respectively. By comparison of amino acid sequences, 13 HA serotypes could be divided into two families, i.e., an H1 group (H1, H2, H5, H6, H8, H9, H11, H12, and H13) and an H3 group (H3, H4, H7, and H10). The relationship was essentially similar to that reported by Air from the comparison of 80 amino-terminal amino acid sequence of 12 HA serotypes (G.M. Air, 1981, Proc. Natl. Acad. Sci. USA 78, 7639-7643). Though a considerable amino acid sequence difference exists between certain HA serotypes, several amino acid residues in fusion peptides (HA2(1-11)) and receptor-binding sites (HA1(98), -134, -138, -153, -183, and -195) were shown to be conserved among the 13 HA serotypes. Human H1 and avian H3, H4, H8, and H10 viruses preferentially bound NeuAc alpha 2,3Gal sequences, whereas human H2 and H3 and avian H6 and H9 viruses bound NeuAc alpha 2,6Gal sequences, although the amino acid residues at position 226 of human H2 and avian H6 and H9 serotype HAs are glutamine. These results show that the amino acid residue at position 226 is not necessarily a determinant of receptor specificity for all serotypes.     

Comparison of genomic and amino acid sequences of eight Japanese encephalitis virus isolates from bats

We compared nucleotide and deduced amino acid sequences of eight Japanese encephalitis virus (JEV) isolates derived from bats in China. We also compared the bat JEV isolates with other JEV isolates available from GenBank to determine their genetic similarity. We found a high genetic homogeneity among the bat JEVs isolated in different geographical areas from various bat species at different time periods. All eight bat JEV isolates belonged to genotype III. The mean evolutionary rate of bat JEV isolates was lower than those of isolates of other origin, but this difference was not statistically significant. Based on these results, we presume that the bat JEV isolates might be evolutionarily conserved. The eight bat JEV isolates were phylogenetically similar to mosquito BN19 and human Liyujie isolates of JEV. These results indicate that bats might be involved in natural cycle of JEV.     

Comparison of complete nucleotide sequences and genome organization of six distinct cherry leaf roll virus isolates from New Zealand

The complete genomic sequences of six phenotypically distinct cherry leaf roll virus (CLRV) isolates from New Zealand were determined and compared. RNA1 and RNA2 are 7,919-7,921 and 6,361-6,363 nucleotides in length, respectively, excluding the poly(A) tails. Both genome segments contain a single open reading frame and encode one polyprotein of 2,113 amino acids for RNA1 and 1,590 aa for RNA2, with corresponding molecular masses of 235.5-236.1 kDa and 174.2-174.8 kDa, respectively. The six RNA1 sequences share 92.0-99.7 % nt and 97.0-99.5 % aa similarity, and the RNA2 sequences share 91.4-99.8 % nt and 94.3-99.7 % aa similarity. Phylogenetic analysis established close associations between the six New Zealand CLRV isolates and members of the subgroup C nepoviruses.     

Complete genome sequences and phylogenetic analysis of hepatitis delta viruses isolated from nine Turkish patients

Hepatitis delta virus (HDV) is a subviral agent of hepatitis B virus (HBV), and its life cycle is dependent on HBV. It is commonly accepted that HDV has eight distinct genotypes. In this study, the complete nucleotide sequences of HDV genomes isolated from nine Turkish patients were obtained by RT-PCR using two pairs of primers that cover the entire HDV genome. PCR products were sequenced directly. The results showed that these 9 isolates were approximately 1680 base pairs in length and clustered in the genotype HDV-1 branch when phylogenetic analysis was done with the sequences together with the complete sequences of HDV genomes representing each genotype retrieved from GenBank. Analysis of a portion of the large hepatitis D antigen (L-HDAg) gene showed that sequence similarity among these Turkish isolates is between 87.4 and 97.1%, and the Turkish isolates have the most sequence similarity to HDV-1 (90.5%), while they have the least sequence similarity to HDV-3 (64.1%). Full-genome analysis indicates that the sequence similarity is between 80.7 and 95.4%, and the highest sequence similarity is 84.8% (between the Turkish isolates and HDV-1). The lowest sequence similarity is 56.4% (between the Turkish isolates and HDV-3). In conclusion, phylogenetic analysis shows that the Turkish HDV isolates belong to HDV-1.     

Nucleotide and deduced amino acid sequences of the glycoprotein gene of rabies virus vaccine strain Vnukovo-32

Summary The glycoprotein gene of the rabies virus vaccine strain Vnukovo-32 was sequenced and the deduced protein sequence was analyzed and compared with that of various laboratory and street strains. The amino acid sequence homologies of strain Vnukovo-32 were compared with fixed strains ERA, SAD B19, PV, HEP-Flury, CVS and two street strains, canine and CXX89-1, were 98.9 %(6 replacements), 98.3% (9), 96.2% (20), 91.4% (45), 87.0% (68), 93.5% (34) and 91.4% (45), respectively. Sequence alignments of the proteins revealed that the most conserved region is the ectodomain, whereas the transmembrane and cytoplasmic domains showed significant divergence.     

Nucleotide sequence of the genome and complete amino acid sequence of the polyprotein of tick-borne encephalitis virus

The sequence of the genome of tick-borne encephalitis (TBE) virus (Far Eastern subtype, strain Sofjin) coding for structural proteins and nonstructural protein NS1 has been previously reported (A. G. Pletnev, V. F. Yamshchikov, and V. M. Blinov, 1986, FEBS Lett. 200, 317-321; Yamshchikov and Pletnev, 1988, Nucleic Acids Res. 16, 7750. Now we have cloned and sequenced the genomic RNA that encodes all nonstructural proteins. Together with our earlier sequence analyses, these data show that the TBE genome is 10,477 bases in length with a single open reading frame extending from nucleotides 127 to 10,363, encoding 3412 amino acids. The 5'- and 3'-noncoding regions have stem-loop structures. The polyprotein precursor is proteolytically cleaved, apparently by a mechanism resembling that proposed for the expression of polyproteins of the other flaviviruses, such as yellow fever and Kunjin viruses. The deduced TBE gene order is 5'-C-pre(M)M-E-NS1-NS2A-NS2B-NS3-ns4a-NS4B -NS5-3'. The genome structure and the polyprotein of TBE virus is similar to mosquito-borne flaviviruses, although TBE virus is transmitted by ticks. Comparison of the sequence homology of polyproteins of flaviviruses suggests that TBE virus is more closely related to yellow fever virus than to other serological subgroups of flaviviruses. The hydrophobicity profile of the TBE polyprotein is similar to those of other flaviviruses. Nonstructural proteins NS2A, NS2B, ns4a, and NS4B are extremely hydrophobic, suggesting that these proteins are likely associated with cellular membranes. Proteins E, NS1, NS3, and NS5 are the most conserved and these proteins may be involved in the general activities related to viral reproduction.     

北京地区TT病毒分离株全基因的克隆及序列测定

目的 克隆和测定北京地区ＴＴ病毒（ＴＴＶ）分离株全基因序列。方法 采用聚合酶链反应（ＰＣＲ）技术从１例临床非甲￣庚型肝炎病人血清中分段扩增ＴＴＶ全基因,获得５个互相重叠的片段,分别长１９９ｂｐ（１－１９９）,１２６７ｂｐ（９０－１３５６,８７８ｂｐ（１３５４－２２３１,１１２９ｂｐ（２１７８－３３０６,４３４ｂｐ（３３０６－３７３９）,用标准的分子生物学方法测定了这５个序列,从而得到全长ＴＴＶ基因     

Characterization of the biological properties and complete genome sequence analysis of a cattle-derived rabies virus isolate from the Guangxi province of southern China

In this study, a street rabies virus isolate, GXHXN, was obtained from the brain of one rabid cattle in Guangxi province of southern China. To characterize the biological properties of GXHXN, we first evaluated its pathogenicity using 4-week-old adult mice. GXHXN was highly pathogenic with a short incubation period and course of disease. Its LD₅₀ of 10^?6.86/mL is significantly higher than the LD₅₀ of 10^?5.19/mL of GXN119, a dog-derived rabies virus isolate. It also displayed a higher neurotropism index than the rRC-HL strain. However, the relative neurotropism index of GXHXN was slightly lower than that of GXN119. Analyzing antigenicity using anti-N and anti-G monoclonal antibodies (MAbs), all tested anti-N MAbs reacted similarly to GXHXN, CVS, and rRC-HL, but the reaction of anti-N MAbs to GXHXN was slightly different from GXN119. Moreover, 2/11 tested anti-G mAbs showed weaker reactivity to GXHXN than rRC-HL, whereas 4/11 showed stronger reactivity to GXHXN than CVS and GXN119, indicating that the structures of G might differ. In order to understand its genetic variation and evolution, the complete GXHXN genome sequence was determined and compared with the known 12 isolates from other mammals. A total of 42 nucleotide substitutions were found in the full-length genome, including 15 non-synonymous mutations. The G gene accounts for the highest nucleotide substitution rate of 0.70 % in ORF and an amino acid substitution rate of 0.95 %. Phylogenetic trees based on the complete genome sequence as well as the N and G gene sequences from 37 known rabies isolates from various mammals demonstrated that the GXHXN is closely related to the BJ2011E isolate from a horse in Beijing, the WH11 isolate from a donkey in Hubei, and isolates from dogs in the Fujian and Zhejiang provinces. These findings will be helpful in exploring the molecular mechanisms underlying interspecies transmission and the genetic variation of the rabies virus in different mammal species.     

The complete nucleotide sequence and genome organization of a novel carmovirus--honeysuckle ringspot virus isolated from honeysuckle

A virus associated with yellow-to-purple ringspot on honeysuckle plants has been detected and tentatively named honeysuckle ringspot virus (HnRSV). The complete nucleotide sequence of HnRSV from infected honeysuckle has been determined. The genomic RNA of HnRSV is 3,956 nucleotides in length and is predicted to contain five open reading frames (ORFs). Comparisons of the amino acid sequences of the ORFs of HnRSV with those of members of the family Tombusviridae show that HnRSV is closely related to members of the genus Carmovirus. Phylogenetic analysis based on the amino acid sequences of RdRp and coat protein and nucleotide sequences of the whole genome revealed that HnRSV forms a subgroup with the carmoviruses. Together, our results support the classification of HnRSV as a member of a new species in the genus Carmovirus, family Tombusviridae.     

Characterization of the complete genome of ribgrass mosaic virus isolated from Plantago major L. from New Zealand and Actinidia spp. from China

The complete genomes of tobamovirus isolates from Plantago major L. from New Zealand (NZ-439), Plantago sp. from Germany (Kons 1105), Actinidia chinensis (Actinidia-AC) and A. deliciosa (Actinidia-AD) from China were sequenced and compared to previously published tobamovirus genomes. Their genome organization and phylogenetic analysis of the putative replicase component, replicase readthrough component, movement protein, coat protein and complete genome placed all four isolates in subgroup 3 of the tobamoviruses. The complete genomes differed from each other by <8.5% and from published sequences of turnip vein clearing virus and youcai mosaic virus by about 12-13% and 19-20%, respectively. The aa sequences of the individual ORFs of the Plantago and Actinidia isolates differed from each other by <4% and were most similar to published (partial) sequences of ribgrass mosaic virus (RMV). We propose that these sequences constitute the first complete published sequences for RMV.     

Analysis of the complete genome of a virus associated with twisted leaf disease of cherry reveals evidence of a close relationship to unassigned viruses in the family Betaflexiviridae

The genome of a virus associated with cherry twisted leaf disease (CTLaV, isolate ZH) was sequenced and consists of 8431 nucleotides, excluding a poly(A) tail at the 3′ end. Genome analysis shows that CTLaV-ZH represents a new and distinct species and has a genome organization similar to those of unassigned viruses in the family Betaflexiviridae. The CTLaV-ZH genome has five open reading frames (ORFs), with putative ORFs within ORF2 and ORF5, identified as ORF2a and ORF5a, respectively. The AUG start codons of ORF2a and ORF5a are in contexts suitable for efficient translation, with appropriate stop codons in frame.     

FBR murine osteosarcoma virus. II. Nucleotide sequence of the provirus reveals that the genome contains sequences acquired from two cellular genes

The complete nucleotide sequence of the FBR proviral DNA has been determined. The provirus of 3791 nucleotides (specifying a genome of 3284 bases) encodes a single gag- fos fusion product of 554 amino acids. The fos portion of the gene lacks the sequences which code for the first 24 and the last 98 amino acids of the 380-amino acid mouse c- fos gene product. In addition, the coding region has sustained three in-frame deletions, one in the p30gag portion, and two in the fos region, as compared to the sequences of AKR-MLV and the c- fos gene, respectively. The gene product terminates in sequences, termed v-fox, that are present in uninfected mouse DNA at loci unrelated to the c- fos gene. The c-fox gene(s) is expressed as an abundant class of polyadenylated RNA in normal mouse tissues.     

中国分离乙脑病毒与灭活疫苗株(P3株)E基因差异分析

目的 分析我国近年来从蚊虫及患者标本中分离的乙脑病毒与灭活疫苗株（P3株）之间在E基因区段核苷酸及氨基酸差异。方法 从GenBank中获取相应乙脑病毒株E基因区段核苷酸序列，通过Clustal X（1．8）、DNASTAR、GENEDOC（3．2）等生物学软件进行分析。结果 P3株与福建分离株之间核苷酸同源性在98．3％-98．5％之间、氨基酸同源性在98．2％-98．6％之间；P3株与上海分离株之间核苷酸同源性在88．0％-88．5％之间、氨基酸同源性在98．0％-98．4％之间。E基因区段500个氨基酸中P3株与所有新分离乙脑病毒株之间共存在19个位点的差异，其中在E-76、E-306、E-408处所有新分离毒株与P3株存在共同的差异；在E-160、E-487处福建分离株与P3株存在共同差异；上海分离株与P3株在E-129、E-222、E-227、E-366存在共同差异。结论 上海蚊虫中分离的Ⅰ型乙脑病毒和福建省脑炎患者中分离的Ⅲ型乙脑病毒与P3株在E基因的部分氨基酸位点存在差异，但均不处在影响病毒生物学特性的关键位点。     

Nucleotide and deduced amino acid sequences of genes encoding the structural and nonstructural NS1 proteins of a dengue-2 virus isolated in China

We have determined the nucleotide and encoded amino acid sequences of the capsid, membrane precursor, membrane, envelope, and nonstructural NS1 protein genes of a dengue-2 virus (D2-04) isolated from a patient in Hainan, China. The sequenced region contains a gene organization similar to that of other flaviviruses. The overall amino acid sequence similarity between D2-04 and other dengue-2 viruses is greater than 92%, whereas that between D2-04 and members of the other dengue serotypes is about 65%.The nucleotide sequence data reported in this paper have been deposited in the EMBL database under the following accession numbers: X 65239 for the capsid (C), X 65240 for the envelope (E), X 65241 for the nonstructural (NS1), and X 65242 for the premembrane/membrane (prM/M) protein genes.     

Variation of the nucleotide and encoded amino acid sequences of the envelope gene from eight dengue-2 viruses

Summary The nucleotide sequences of the envelope genes from five Thai and three Sri Lankan dengue-2 viruses were determined by sequencing the viral RNA using synthetic oligonucleotide primers. The results were compared with the four published dengue-2 envelope sequences to obtain a classification of these viruses, which showed that the Thai isolates could be divided into two separate groups while the Sri Lankan isolates were distinct. There was no correlation between disease severity and envelope protein sequence, or between year of isolation and sequence. No particular amino acid changes were associated with virulence or a change in hydrophilic region which could perhaps act as an epitope.