scholarly journals Molecular characterization of the major capsid protein VP6 of bovine group B rotavirus and its use in seroepidemiology

2005 ◽  
Vol 86 (9) ◽  
pp. 2569-2575 ◽  
Author(s):  
Hiroshi Tsunemitsu ◽  
Mariko Kamiyama ◽  
Kenji Kawashima ◽  
Ken Katsuda ◽  
Mariko Kohmoto ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Expression System ◽  
Baculovirus Expression System ◽  
Reading Frame ◽  
High Prevalence ◽  
Group A Rotaviruses ◽  
Group A ◽  
Vp6 Gene ◽  
Group B

The major inner capsid protein (VP6) gene of the bovine group B rotavirus (GBR) Nemuro strain is 1269 nt in length and contains one open reading frame encoding 391 aa. Nucleotide and amino acid sequence identities of the Nemuro VP6 gene compared with the published corresponding human and rodent GBR genes were respectively 66–67 and 70–72 %, which are notably lower than those between human and rodent viruses (72–73 and 83–84 %, respectively). Overall identities of VP6 genes among GBRs were substantially lower than those among both group A rotaviruses (GARs) and group C rotaviruses (GCRs) derived from different species of mammals. These results demonstrate that bovine GBR is remarkably distinct from other GBRs and that GBRs from different species may have had a longer period of divergence than GARs and GCRs. Recombinant VP6 was generated with a baculovirus expression system and used for an ELISA to detect GBR antibodies. All 13 paired sera from adult cows with GBR-induced diarrhoea in the field showed antibody responses in the ELISA. In serological surveys of GBR infection using the ELISA, 47 % of cattle sera were positive for GBR antibodies, with a higher antibody prevalence in adults than in young cattle. In pigs, a high prevalence of GBR antibodies (97 %) was detected in sera from sows. These results suggest that GBR infection is common in cattle and pigs, notwithstanding the scarcity of reports of GBR detection in these species to date.

scholarly journals Molecular Characterization of a Subgroup Specificity Associated with the Rotavirus Inner Capsid Protein VP2

2008 ◽  
Vol 82 (6) ◽  
pp. 2752-2764 ◽  
Author(s):  
Sarah M. McDonald ◽  
John T. Patton
Keyword(s):  
Monoclonal Antibody ◽  
Capsid Protein ◽  
Neutralizing Antibodies ◽  
Bovine Rotavirus ◽  
Single Region ◽  
Molecular Determinants ◽  
High Resolution Structure ◽  
Group A Rotaviruses ◽  
Group A

ABSTRACT Group A rotaviruses are classified into serotypes, based on the reactivity pattern of neutralizing antibodies to VP4 and VP7, as well as into subgroups (SGs), based on non-neutralizing antibodies directed against VP6. The inner capsid protein (VP2) has also been described as a SG antigen; however, little is known regarding the molecular determinants of VP2 SG specificity. In this study, we characterize VP2 SGs by correlating genetic markers with the immunoreactivity of the SG-specific monoclonal antibody (YO-60). Our results show that VP2 proteins similar in sequence to that of the prototypic human strain Wa are recognized by YO-60, classifying them as VP2 SG-II. In contrast, proteins not bound by YO-60 are similar to those of human strains DS-1 or AU-1 and represent VP2 SG-I. Using a mutagenesis approach, we identified residues that determine recognition by either YO-60 or the group A-specific VP2 monoclonal antibody (6E8). We found that YO-60 binds to a conformationally dependent epitope that includes Wa VP2 residue M328. The epitope for 6E8 is also contingent upon VP2 conformation and resides within a single region of the protein (Wa VP2 residues A440 to T530). Using a high-resolution structure of bovine rotavirus double-layered particles, we predicted these epitopes to be spatially distinct from each other and located on opposite surfaces of VP2. This study reveals the extent of genetic variation among group A rotavirus VP2 proteins and illuminates the molecular basis for a previously described SG specificity associated with the rotavirus inner capsid protein.

scholarly journals The Genome Segments of a Group D Rotavirus Possess Group A-Like Conserved Termini but Encode Group-Specific Proteins

2010 ◽  
Vol 84 (19) ◽  
pp. 10254-10265 ◽  
Author(s):  
Eva Trojnar ◽  
Peter Otto ◽  
Bernhard Roth ◽  
Jochen Reetz ◽  
Reimar Johne
Keyword(s):  
Sequence Data ◽  
Open Reading Frames ◽  
Degenerate Primers ◽  
Nucleotide Sequence Data ◽  
Reading Frame ◽  
Group A Rotaviruses ◽  
Group A ◽  
Specific Proteins ◽  
Group B ◽  
Group D

ABSTRACT Rotaviruses are a leading cause of viral acute gastroenteritis in humans and animals. They are grouped according to gene composition and antigenicity of VP6. Whereas group A, B, and C rotaviruses are found in humans and animals, group D rotaviruses have been exclusively detected in birds. Despite their broad distribution among chickens, no nucleotide sequence data exist so far. Here, the first complete genome sequence of a group D rotavirus (strain 05V0049) is presented, which was amplified using sequence-independent amplification strategies and degenerate primers. Open reading frames encoding homologues of rotavirus proteins VP1 to VP4, VP6, VP7, and NSP1 to NSP5 were identified. Amino acid sequence identities between the group D rotavirus and the group A, B, and C rotaviruses varied between 12.3% and 51.7%, 11.0% and 23.1%, and 9.5% and 46.9%, respectively. Segment 10 of the group D rotavirus has an additional open reading frame. Generally, phylogenetic analysis indicated a common evolution of group A, C, and D rotaviruses, separate from that of group B. However, the NSP4 sequence of group C has only very low identities in comparison with cogent sequences of all other groups. The avian group A NSP1 sequences are more closely related to those of group D than those of mammalian group A rotaviruses. Most interestingly, the nucleotide sequences at the termini of the 11 genome segments are identical between group D and group A rotaviruses. Further investigations should clarify whether these conserved structures allow an exchange of genome segments between group A and group D rotaviruses.

scholarly journals Molecular characterization of unusual G10P[33], G6P[14] genomic constellations and evidence of zooanthroponosis in bovines

2020 ◽  
Author(s):  
P.M Sawant ◽  
S Digraskar ◽  
V. Gopalkrishna
Keyword(s):  
Western India ◽  
Prevention Strategies ◽  
Species Barrier ◽  
Genetic Characteristics ◽  
Group A Rotaviruses ◽  
Group A ◽  
Lineage 2 ◽  
Vp6 Gene ◽  
Neonatal Calves

AbstractGroup A rotaviruses (RVA) are a major cause of diarrhea in neonatal calves and children. The present study examined G/P combinations and genetic characteristics of RVAs in diarrheic bovine calves in Western India. RVAs were detected in 27 samples (17.64%) with predominance of G10P[11] (51.85%), followed by previously unreported genomic constellations, G6P[14] (14.81%), and, G6P[4] (7.40%) and G10P[33] (3.70%). Sequencing and phylogenetic analysis revealed circulation of G10 (Lineage-5), G6 (Lineage-2), P[11] (Lineage-3), P[14] (proposed Lineage-8) and P[4] (Lineage-3) genotypes. The predominant G10P[11] strains were typical bovine strains and exhibited genotypic homogeneity. The rare, G10P[33] strain, had VP7 and VP4 genes of bovine origin but resemblance of VP6 gene with simian strain indicated possible reassortment between bovine and simian (SA11-like) strains. The VP6 and VP7 genes of other two rare strains, G6P[14] and G6P[4], were similar to those of bovine stains, but the VP4 was closely related to those of the human-bovine like and human strains, respectively. Additionally, in VP4 gene phylogenetic tree Indian P[14] strains constituted a closely related genetic cluster distinct from the other P[14] strains, hence Lineage-8 was proposed for them. These findings indicated that bovines could serve as source for anthropozoonotic transmission of G6P[14] strains while zooanthroponotic transmission followed by reassortment with human strain gave rise to G6P[4] strains. The observations of present study reinforce the potential of rotaviruses to cross the host-species barrier and undergo reassortant to increase genetic diversity which necessitates their continuous surveillance for development and optimization of prevention strategies against zoonotic RVAs.

Expression, Purification, and Characterization of Recombinant Human α1-Antitrypsin Produced Using Silkworm–Baculovirus Expression System

2018 ◽  
Vol 60 (12) ◽  
pp. 924-934 ◽  
Author(s):  
Yoshiki Morifuji ◽  
Jian Xu ◽  
Noriko Karasaki ◽  
Kazuhiro Iiyama ◽  
Daisuke Morokuma ◽  
...  
Keyword(s):  
Expression System ◽  
Baculovirus Expression System ◽  
Purification And Characterization ◽  
Baculovirus Expression

Cloning and functional characterization of an uncoupling protein homolog in hummingbirds

2001 ◽  
Vol 5 (3) ◽  
pp. 137-145 ◽  
Author(s):  
CLAUDIA R. VIANNA ◽  
THILO HAGEN ◽  
CHEN-YU ZHANG ◽  
ERIC BACHMAN ◽  
OLIVIER BOSS ◽  
...  
Keyword(s):  
Uncoupling Protein ◽  
Expression System ◽  
Functional Characterization ◽  
Pectoral Muscle ◽  
Mitochondrial Fraction ◽  
Mrna Levels ◽  
Coding Region ◽  
Reading Frame ◽  
Rapid Amplification

The cDNA of an uncoupling protein (UCP) homolog has been cloned from the swallow-tailed hummingbird, Eupetomena macroura. The hummingbird uncoupling protein (HmUCP) cDNA was amplified from pectoral muscle (flight muscle) using RT-PCR and primers for conserved domains of various known UCP homologs. The rapid amplification of cDNA ends (RACE) method was used to complete the cloning of the 5′ and 3′ ends of the open reading frame. The HmUCP coding region contains 915 nucleotides, and the deduced protein sequence consists of 304 amino acids, being ∼72, 70, and 55% identical to human UCP3, UCP2, and UCP1, respectively. The uncoupling activity of this novel protein was characterized in yeast. In this expression system, the 12CA5-tagged HmUCP fusion protein was detected by Western blot in the enriched mitochondrial fraction. Similarly to rat UCP1, HmUCP decreased the mitochondrial membrane potential as measured in whole yeast by uptake of the fluorescent potential-sensitive dye 3′,3-dihexyloxacarbocyanine iodide. The HmUCP mRNA is primarily expressed in skeletal muscle, but high levels can also be detected in heart and liver, as assessed by Northern blot analysis. Lowering the room’s temperature to 12–14°C triggered the cycle torpor/rewarming, typical of hummingbirds. Both in the pectoral muscle and heart, HmUCP mRNA levels were 1.5- to 3.4-fold higher during torpor. In conclusion, this is the first report of an UCP homolog in birds. The data indicate that HmUCP has the potential to function as an UCP and could play a thermogenic role during rewarming.

Molecular characterization of the first human G15 rotavirus strain of zoonotic origin from the bovine species

2021 ◽  
Vol 102 (4) ◽  
Author(s):  
Takeshi Tsugawa ◽  
Yoshiki Fujii ◽  
Yusuke Akane ◽  
Saho Honjo ◽  
Kenji Kondo ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Molecular Characterization ◽  
Human Infection ◽  
Potential Reservoir ◽  
Group A Rotaviruses ◽  
Paediatric Outpatient ◽  
Group A ◽  
Human Infections ◽  
Bovine Species

Group A rotaviruses (RVAs) infect a wide variety of mammalian and avian species. Animals act as a potential reservoir to RVA human infections by direct virion transmission or by contributing genes to reassortants. Here, we report the molecular characterization of a rare human RVA strain Ni17-46 with a genotype G15P[14], isolated in Japan in 2017 during rotavirus surveillance in a paediatric outpatient clinic. The genome constellation of this strain was G15-P[14]-I2-R2-C2-M2-A13-N2-T9-E2-H3. This is the first report of an RVA with G15 genotype in humans, and sequencing and phylogenetic analysis results suggest that human infection with this strain has zoonotic origin from the bovine species. Given the fact that this strain was isolated from a patient with gastroenteritis and dehydration symptoms, we must take into account the virulence of this strain in humans.

scholarly journals Dominance of the ON1 Genotype of RSV-A and BA9 Genotype of RSV-B in Respiratory Cases from Jeddah, Saudi Arabia

Genes ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 1323
Author(s):  
Hessa A. Al-Sharif ◽  
Sherif A. El-Kafrawy ◽  
Jehad M. Yousef ◽  
Taha A. Kumosani ◽  
Mohammad A. Kamal ◽  
...  
Keyword(s):  
Saudi Arabia ◽  
Respiratory Infections ◽  
Hypervariable Region ◽  
Genotype Distribution ◽  
Potential Development ◽  
The Usa ◽  
Group A ◽  
Syncytial Virus ◽  
Group B

Human respiratory syncytial virus (HRSV) is a main cause of hospital admission for lower respiratory tract infection. In previous studies from Saudi Arabia, higher prevalence of the NA1 genotype in group A was observed from Riyadh and Taif. This study recruited respiratory cases from Jeddah during January to December, 2017. RSV represented 13.4% in the recruited cases with 64% of them belonging to group A and 36% to group B. All group A cases in this study were ON1 type characterized by duplication of 72 nucleotides, 24 amino acids in the C-terminal in the second hypervariable region of the G gene. In addition, for group B all of the cases were clustered under BA9, which had uniquely characterized as duplication of 60 nucleotides in the G protein. Our sequences showed similarity with earlier sequences from Saudi Arabia, Kuwait, Thailand, South Africa, Spain, the USA and Cyprus. Some amino acid substitutions in the investigated sequences would cause a change in potential O-glycosylation and N-glycosylation profiles from prototype ON1. The predominance of the ON1 and BA9 genotype of RSV-A in Jeddah compared to previous Saudi studies showing predominance of the NA1 genotype for group A. This difference in genotype prevalence could be due to fast spread of the ON1 genotype worldwide or due to the flux of travelers through Jeddah during hajj/umrah compared to Riyadh and Taif. This shift in genotype distribution requires continuous surveillance for genetic characterization of circulating respiratory infections including RSV. These findings may contribute to the understanding of RSV evolution and to the potential development of a vaccine against RSV.

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