المؤلفون: Ahmed Hadidi, L. G. Nemchinov, T. D. Verderevskaya, E. Z. Zemtchik, A. M. Shamloul

المصدر: Archives of virology. 145(9)

مصطلحات موضوعية: food.ingredient, Blotting, Western, Molecular Sequence Data, Genome, Viral, Molecular cloning, Biology, Genome, Foveavirus, Virus, Plant Viruses, food, Capsid, Virology, Complementary DNA, Coding region, Amino Acid Sequence, Cloning, Molecular, Rosales, Gene, 3' Untranslated Regions, Phylogeny, Base Sequence, Reverse Transcriptase Polymerase Chain Reaction, Nucleic Acid Hybridization, General Medicine, biology.organism_classification, Apple stem pitting virus, Virus Latency, 5' Untranslated Regions, Sequence Alignment

الوصف: Extraction of viral double-stranded RNA from peach leaves infected with Apricot latent virus (ALV) followed by molecular cloning of synthesized cDNA and its sequencing, suggested that ALV is a new virus, whose coat protein (CP) coding region contains Apple stem pitting virus (ASPV)-related sequences. The sequenced portion of the ALV genome (1,444 nt) includes the putative CP gene and the 3' non-translated region. The 5' portion of this fragment (1-651 nt) is highly distinct whereas the 3' portion is 77% identical to the corresponding region of ASPV. Molecular hybridization experiments using a cRNA probe to ASPV with ALV-infected leaf tissue extracts also revealed that the genome of ALV contains nucleotide sequences related to that of ASPV. Western blots of tissue extracts indicated that ALV coat protein reacted with polyclonal antiserum against ASPV; however, the ALV CP differs in size from that of ASPV. ALV was graft-transmitted to several Prunus rootstocks. Based on the available sequence data, serological observations and bioassays we propose that ALV is a new species in the genus Foveavirus, typified by ASPV. ALV-specific PCR-primers and viral-specific cRNA probes developed in this investigation may be useful for detecting the virus and for studying its epidemiology and geographical distribution.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::baec1f92110d72afad40590a812a7929Test
https://pubmed.ncbi.nlm.nih.gov/11043942Test

المؤلفون: A. Kitaoka, Hajime Mori, K. Kudo, H. Sato, S. Hara, Kaeko Kamei, Y. Inui, Kazuhiro Nakajima, T. Kamata, R. Takano, E. Emoto

المصدر: Archives of virology. 144(7)

مصطلحات موضوعية: viruses, Recombinant Fusion Proteins, Molecular Sequence Data, Molecular cloning, Biology, Recombinant virus, Epitope, law.invention, Mice, Open Reading Frames, Capsid, law, Virology, Complementary DNA, Polyhedrin, Birnaviridae, Coding region, Animals, Amino Acid Sequence, Cloning, Molecular, fungi, Fishes, virus diseases, Ascites, General Medicine, biochemical phenomena, metabolism, and nutrition, Bombyx, Molecular biology, Fusion protein, Molecular Weight, Recombinant DNA, Capsid Proteins

الوصف: cDNA of yellowtail ascites virus (YAV) segment A encoding a polyprotein of VP2, NS, and VP3 has been cloned. Comparison of the nucleotide and the deduced amino acid sequences showed very high homology between YAV and other aquatic birnaviruses. The two small open reading frames (VP5) besides the 5′ terminus of the VP2 gene were found on segment A of YAV. Proteins encoded by cDNAs from segment A and the serotype-specific epitope region on VP2 were expressed using a baculovirus vector. Western blot analysis confirmed that a polyprotein was expressed and processed into VP2 and VP3 in insect cells infected with the recombinant baculovirus containing the complete polyprotein coding region. In the case of expression in silkworm larvae, only VP3 was detected in hemocytes and fat body of silkworm larvae infected with the recombinant virus. The recombinant fusion protein consisting of VP2 epitope region and polyhedrin was expressed in insect cells and cross-reacted with a mouse monoclonal antibody against VP2 which had a neutralizing activity to YAV.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::44350bd41bd4dbe8ecaeb8cbe730a180Test
https://pubmed.ncbi.nlm.nih.gov/10481746Test

المؤلفون: Wendy Werner, Richard J. McCoy, Michael Sheppard, Michael A. Johnson

المصدر: Archives of virology. 143(3)

مصطلحات موضوعية: viruses, TATA box, RNA Splicing, Molecular Sequence Data, Genome, Viral, Molecular cloning, Biology, Upstream Stimulatory Factor, Birds, chemistry.chemical_compound, Plasmid, Virology, Complementary DNA, Coding region, Animals, Humans, RNA, Messenger, Cloning, Molecular, Promoter Regions, Genetic, Gene, Cells, Cultured, Genetics, Base Sequence, Aviadenovirus, Chromosome Mapping, General Medicine, Molecular biology, chemistry, DNA, Viral, Chickens, DNA

الوصف: The region of the fowl adenovirus serotype 10 (FAV-10) genome containing the major late promoter (MLP) and leader sequences was determined and appropriate genomic fragments were cloned and sequenced. A TATA box was identified and the location of the putative transcription start site was determined. By using synthetic primers from the transcription start site in conjunction with oligonucleotides from the coding regions of the penton base and hexon genes, cDNA was produced from late mRNA isolated from cell cultures infected with FAV-10 at 24 h post-infection. The resulting cDNA was cloned and sequenced and the leader sequences thus identified. It was found that the FAV-10 MLP utilized only two leader sequences (a bipartite leader). By comparison with human adenoviruses (HAVs) it appeared that the second leader in HAVs was absent from the FAV-10. The second leader sequences of FAV-10 was larger than either the second or third leaders of HAVs, but was 29 basepairs shorter than the combined size of the leader sequences 2 and 3 from HAV-2. To confirm the transcription start site and leader sequences, single stranded cDNA was produced from mRNA using the primers from within the coding sequence for the penton base or hexon. A tail of dGTP's was added and cDNA synthesis was completed using an oligonucleotide from within the hexon or penton base coding sequence and a second poly-dCTP oligonucleotide. Sequencing of the resultant G-tailed DNA confirmed the location of the transcription start site as an adenosine residue 24 basepairs upstream from the 3-prime (3') end of the TATA box. Sequencing 5' of the TATA box failed to reveal any sequence similarity with the human adenovirus upstream stimulatory factor (USF). Various plasmids were constructed which placed the determined sequences of the MLP, leader, and the region upstream of the TATA box linked to the co-acetyl acid transferase (CAT) gene. These expression plasmids in transient expression assays of CAT activity in primary chicken kidney cell culture with or without FAV-10 co-infection were determined. These experiments showed that the cassette containing sequences 5' of the TATA box expressed CAT to a much greater level than cassettes not containing this upstream region and that the presence of virus significantly increased the activity of the promoter following the onset of viral DNA replication. Without the 5' region, cassettes failed to express above background levels. These results suggest that the basic structure of the fowl adenovirus MLP is similar to that of the human adenovirus although it utilizes a bipartite rather than a tripartite leader sequence.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::57fadc393ad15f67aaa3722b9dd2c1ccTest
https://pubmed.ncbi.nlm.nih.gov/9572553Test