The research of p53 is being conducted to find the mechanisms of tumorigenesis and to treat various cancers. Homeodomain-interacting protein kinase2 (HIPK2) is an important factor to regulate p53 and to increase the stability of p53. Activation of HIPK2 leads to the selective phosphorylation of p53, resulting in growth arrest and the enhancement of apoptosis. In this study, the canine HIPK2 cDNA fragments were obtained, and their overlapping regions were aligned to give a total sequence of 3489 bp. The canine HIPK2 cDNA (GenBank accession number; AY800385) shares 93% and 90% sequence identity with those of human and mouse HIPK2, respectively. The canine HIPK2 cDNA contains an open reading frame encoding 1163 amino acid residues and the predicted amino acid sequence has 98% and 96% identity with those of human and mouse, respectively. The deduced amino acid sequence of canine HIPK2 has also all domains' sites compared with human and mouse HIPK2. Therefore, these structural similarities suggested that the canine HIPK2 shares the basic biological functions that HIPK2 exhibit in other species.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary/chemistry/genetics ; Dogs/metabolism/*physiology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; Protein-Serine-Threonine Kinases/*genetics ; Sequence Alignment ; Sequence Analysis, DNA

Twelve Korean infectious bronchitis viruses (IBVs) were isolated in the field from chickens suspected of being carriers of infectious bronchitis between 2001 and 2003. The S1 glycoprotein genes of these IBV isolates were amplified by reverse transcriptase-polymerase chain reaction (RTPCR) and analyzed by restriction fragment length polymorphism (RFLP) analysis. These Korean IBV isolates were classified into three groups according to their RFLP patterns obtained using the restriction enzyme HaeIII. Half of the twelve isolates were similar to the KM91 RFLP pattern, which is a common pattern in Korea. Three more isolates were related to the Arkansas strain pattern, but with some unique variations. The other three viruses showed variant RFLP patterns. For a comparison with the published sequences for non-Korean IBV strains, amplified PCR products from the twelve isolates were cloned and sequenced. The Korean IBV field isolates had 71.2-99.7% nucleotide sequence homology with each other and 45.9-80.7% nucleotide sequence homology with non-Korean IBV strains. With respect to the deduced amino acid sequence, the Korean IBV isolates had 71.5-99.3% similarity with each other and 44.9-80.3% similarity with non-Korean IBV strains. Phylogenetic tree analysis revealed that some of the IBV isolates appear to belong to a new group, different from the non-Korean IBV strains or from previously isolated Korean IBV strains. Specifically, the new Korean IBV isolates K10217-03, K3-3 and K1255-03 represented a separate group. These findings suggest that the Korean IBVs appear to be continuously evolving.
Amino Acid Sequence ; Animals ; Coronavirus Infections/*veterinary/virology ; Glycoproteins/chemistry/*genetics ; Infectious bronchitis virus/*classification/*genetics/isolation & purification ; Molecular Sequence Data ; *Phylogeny ; Polymorphism, Restriction Fragment Length ; Poultry ; Poultry Diseases/*virology ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Alignment/veterinary ; Sequence Analysis ; Sequence Homology, Amino Acid ; Viral Proteins/chemistry/genetics

Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.
Amino Acid Sequence ; Animal Diseases/*virology ; Animals ; Animals, Zoo/*virology ; Coronaviridae Infections/*veterinary/virology ; Coronavirus, Canine/genetics/*isolation & purification ; Fatal Outcome ; Female ; Male ; Molecular Sequence Data ; Sequence Alignment ; Sequence Homology, Amino Acid ; Ursidae/*virology ; Viral Proteins/chemistry

Species identification using DNA sequences is the basis for DNA taxonomy. In this study, we sequenced the ribosomal large-subunit RNA gene sequences (3,037-3,061 bp) in length of 13 Chinese Theileria stocks that were infective to cattle and sheep. The complete 28S rRNA gene is relatively difficult to amplify and its conserved region is not important for phylogenetic study. Therefore, we selected the D2-D3 region from the complete 28S rRNA sequences for phylogenetic analysis. Our analyses of 28S rRNA gene sequences showed that the 28S rRNA was useful as a phylogenetic marker for analyzing the relationships among Theileria spp. in ruminants. In addition, the D2-D3 region was a short segment that could be used instead of the whole 28S rRNA sequence during the phylogenetic analysis of Theileria, and it may be an ideal DNA barcode.
Animals ; Base Sequence ; China ; DNA, Ribosomal/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 28S/genetics ; Ruminants ; Sequence Alignment ; Sequence Analysis, DNA/veterinary ; Theileria/*classification/genetics/isolation & purification ; Theileriasis/*parasitology

Although little is known on the function of gammadelta T lymphocytes, there is increasing evidence that gammadelta T lymphocytes are early responders and modulators of immune responses against pathogens and cytokines such as IL-2, IL-7, IL-15 and TNF-alpha. To study the role TNF-alpha on gammadelta T lymphocytes, we cloned bovine TNF-alpha. Sequence analysis revealed that a new allele form of bovine TNF-alpha was cloned which has 3 additional nucleotide sequences as well as 3 nucleotide substitutions compared with previously reported bovine TNF-alpha. Further studies are needed to document the functional significance of a new allele form of TNF-alpha in cattle.
*Alleles ; Amino Acid Sequence ; Animals ; Base Sequence ; Cattle ; *Cloning, Molecular ; Gene Library ; Molecular Sequence Data ; Receptors, Antigen, T-Cell, gamma-delta/*physiology ; Sequence Alignment/veterinary ; T-Lymphocytes/immunology/*physiology ; Tumor Necrosis Factor-alpha/chemistry/*genetics/physiology

Amplification of the 16S rRNA gene from a blood sample obtained from a dog in southeastern Brazil was used to confirm a naturally acquired Ehrlichia (E.) canis infection. Following isolation and culturing of the new bacterial strain called Uberlandia, partial sequences of the dsb and p28 genes were obtained. The dsb partial sequence of the novel strain was 100% similar to dsb gene sequences of E. canis obtained from different geographic areas around the world. Conversely, the p28 partial sequence for the E. canis Uberlandia strain differed at several nucleotides from other sequences available in GenBank. To confirm the antigenic profile of the Uberlandia strain, an indirect immunofluorescence assay against E. canis antigens was performed using dog sera collected from two different areas in Brazil (Uberlandia and Sao Paulo). The results suggest that both antigens were able to identify animals seropositive for E. canis in Brazil since these Brazilian strains appear to be highly conserved.
Animals ; Antigens, Bacterial/blood/*diagnostic use ; Bacterial Outer Membrane Proteins/genetics/metabolism ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; Brazil ; Dog Diseases/diagnosis/*microbiology ; Dogs ; Ehrlichia canis/*genetics/*immunology/isolation & purification ; Ehrlichiosis/diagnosis/microbiology/*veterinary ; Fluorescent Antibody Technique, Indirect/veterinary ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; RNA, Ribosomal, 16S/genetics/metabolism ; Sequence Alignment/veterinary

From May to June 2002, a total of 16 foot-and mouth disease (FMD) outbreaks due to the serotype O virus, Pan Asia strain, were recorded in Korea. The viruses were identified by antigen ELISA, RT-PCR and sequence analysis. The overall nucleotide sequence divergence of the VP1 region among the 4 isolates in 2002 was 0 to 1.4%, but between O/SKR/2002 and O/SKR/2000 isolates was 1.9-4.9%. Phylogenetic analysis with the some known strains from East Asian countries showed that the 4 Korean isolates in 2002 formed one distinct cluster, which different from clusters of Korean isolates in 2000, with in the same lineage of the ME-SA topotype strains. Deduced amino acid sequences around neutralizable antigenic site on VP1 site of O/SKR/2002 isolates were aligned and compared with other strains. At the antigenic site 1, the replacements of the critical amino acid residues at position 144 from V to L and at position 152 from A to T were observed in O/SKR/2002 viruses. For antigenic site 2 and 4, there were not significant variations in general. At the antigenic site 3, the substitutions of amino acid residues were present at positions 54 and 56 in O/SKR/2002 isolates and an alternative residue I at position 54 are observed only at the sequence of O/SKR/AS/2002 (cow) virus. And the substitution (L-->P) of significant residue at position 144 was detected at the amino acid sequence of the O/SKR/2002 (cow) virus.
Amino Acid Sequence ; Animals ; Antibodies, Viral/blood ; Base Sequence ; Capsid Proteins/genetics/*immunology ; Cattle ; Cattle Diseases/epidemiology/*virology ; Cluster Analysis ; Disease Outbreaks/*veterinary ; Enzyme-Linked Immunosorbent Assay/veterinary ; Epitopes/analysis ; Foot-and-Mouth Disease/epidemiology/*virology ; Foot-and-Mouth Disease Virus/genetics/*immunology ; Korea/epidemiology ; Molecular Sequence Data ; Phylogeny ; RNA, Viral/chemistry/genetics ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Alignment ; Swine ; Swine Diseases/epidemiology/*virology

Avian metapneumovirus (aMPV) causes upper respiratory tract infections in chickens and turkeys. Although the swollen head syndrome (SHS) associated with aMPV in chickens has been reported in Korea since 1992, this is the study isolating aMPV from chickens in this country. We examined 780 oropharyngeal swab or nasal turbinate samples collected from 130 chicken flocks to investigate the prevalence of aMPV and to isolate aMPV from chickens from 2004-2008. Twelve aMPV subtype A and 13 subtype B strains were detected from clinical samples by the aMPV subtype A and B multiplex real-time reverse transcription polymerase chain reaction (RRT-PCR). Partial sequence analysis of the G glycoprotein gene confirmed that the detected aMPVs belonged to subtypes A and B. Two aMPVs subtype A out of the 25 detected aMPVs were isolated by Vero cell passage. In animal experiments with an aMPV isolate, viral RNA was detected in nasal discharge, although no clinical signs of SHS were observed in chickens. In contrast to chickens, turkeys showed severe nasal discharge and a relatively higher titer of viral excretion than chickens. Here, we reveal the co-circulation of aMPV subtypes A and B, and isolate aMPVs from chicken flocks in Korea.
Animals ; Antibodies, Viral/blood ; Base Sequence ; *Chickens ; Glycoproteins/chemistry/genetics ; Metapneumovirus/immunology/*isolation & purification ; Molecular Sequence Data ; Paramyxoviridae Infections/immunology/*veterinary/virology ; *Phylogeny ; Poultry Diseases/immunology/*virology ; RNA, Viral/chemistry/genetics ; Respiratory Tract Infections/immunology/*veterinary/virology ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Alignment ; Sequence Analysis, DNA ; Serotyping ; Specific Pathogen-Free Organisms ; Turkeys

To recognize the molecular biology character, phylogenetic relationship and the state quo prevalent of Canine parvovirus (CPV), Faecal samnples from pet dogs with acute enteritis in the cities of Beijing, Wuhan, and Nanjing were collected and tested for CPV by PCR and other assay between 2006 and 2008. There was no CPV to FPV (MEV) variation by PCR-RFLP analysis in all samples. The complete ORFs of VP2 genes were obtained by PCR from 15 clinical CPVs and 2 CPV vaccine strains. All amplicons were cloned and sequenced. Analysis of the VP2 sequences showed that clinical CPVs both belong to CPV-2a subtype, and could be classified into a new cluster by amino acids contrasting which contains Tyr-->Ile (324) mutation. Besides the 2 CPV vaccine strains belong to CPV-2 subtype, and both of them have scattered variation in amino acids residues of VP2 protein. Construction of the phylogenetic tree based on CPV VP2 sequence showed these 15 CPV clinical strains were in close relationship with Korea strain K001 than CPV-2a isolates in other countries at early time, It is indicated that the canine parvovirus genetic variation was associated with location and time in some degree. The survey of CPV capsid protein VP2 gene provided the useful information for the identification of CPV types and understanding of their genetic relationship.
Amino Acid Sequence ; Animals ; China ; Dog Diseases ; virology ; Dogs ; Genetic Variation ; Molecular Sequence Data ; Parvoviridae Infections ; veterinary ; virology ; Parvovirus, Canine ; chemistry ; classification ; genetics ; isolation & purification ; Phylogeny ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics

Tissue samples of Fabricius' bursa collected from Nanning, Yulin, Beihai and Wuzhou in the provinces of Guangxi in China during the years of 2000-2007, were detected by a established reverse transcriptase polymerase chain reaction (RT-PCR) technique for IBDV. Viral isolation was performed on the positive samples by chicken embryo inoculation via chorio-allantoic membrane (CAM). Results showed that 27 isolates of IBDV were obtained. A set of primers were designed to amplify the vVP2 of 27 isolates by RT-PCR and the PCR products were sequenced. The sequences of all the isolates and reference viruses were analyzed and compared, and their phylogenetic trees were constructed based on the nucleotide sequences. The results indicated that isolate BH11, TZ(3), 050222, YL051, NN0603, NN0611and QX0602 etc, altogether 17 isolates, which accounted for 62.96 percent of total isolates, were identified to be very virulent IBDV (vvIBDV) and have the highest homology to vvIBDV reference strains. In the phylogenetic analysis, they are divided into 3 groups and have a long distance to commonly used vaccine stains. Isolate NN040124 and YL052 were identified as intermediate-plus virulent strains and showed a highest homology to classical strains of 52-70 and STC. 8 isolates of YLZF2, 040131 etc were identified as attenuated vaccine strains and showed a highest homology to classical strain of CU1. The results from the study demonstrated that the viruses prevailing in chickens in these 4 regions in Guangxi province in the recently 7 years were vvIBDV and their origins were complex. The antigenicity of some isolates may have been drifted.
Amino Acid Sequence ; Animals ; Birnaviridae Infections ; epidemiology ; veterinary ; virology ; Chickens ; China ; epidemiology ; Infectious bursal disease virus ; chemistry ; classification ; genetics ; isolation & purification ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Poultry Diseases ; epidemiology ; virology ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics