1.Progress in research of norovirus.
Jia-Dong WANG ; Jun FANG ; Xiao-Hui HAN
Chinese Journal of Virology 2008;24(5):409-413
2.Genome structure and variation of Reynoutria japonica Houtt. chloroplast genome.
Mengtao SUN ; Junxin ZHANG ; Tiran HUANG ; Mingfeng YANG ; Lanqing MA ; Liusheng DUAN
Chinese Journal of Biotechnology 2022;38(5):1953-1964
Reynoutria japonica Houtt., belonging to Polygoneae of Polygonaceae, is a Chinese medicinal herb with the functions of draining dampness and relieving jaundice, clearing heat and detoxifying, dispersing blood stasis and relieving pain, and relieving cough and resolving phlegm. In this study, we carried out high-throughput sequencing for the chloroplast genome sequences of five cultivars of R. japonica and analyzed the genome structure and variations. The chloroplast genomes of the five R. japonica cultivars had two sizes (163 376 bp and 163 371 bp) and a typical circular tetrad structure composed of a large single-copy (LSC) region of 85 784 bp, a small single-copy (SSC) region of 18 616 bp, and a pair of inverted repeat (IR) regions (IRa/IRb) which are spaced apart. A total of 161 genes were obtained by annotation, which consisted of 106 protein-coding genes, 10 rRNA-coding genes, and 45 tRNA-coding genes. The total GC content was 36.7%. Specifically, the GC content in the LSC, SSC, and IR regions were 34.8%, 30.7%, and 42.7%, respectively. Comparison of the whole chloroplast genome among the five cultivars showed that trnk-UUU, rpoC1, petD, rpl16, ndhA, and rpl12 in coding regions had sequence variations. In the phylogenetic tree constructed for the 11 samples of Polygoneae, the five cultivars of R. japonica clustered into one clade near the root and was a sister group of Fallopia multiflora (Thunb.).
Base Composition
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Genome, Chloroplast/genetics*
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Open Reading Frames
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Phylogeny
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Reynoutria
3.Micropeptides: origins, identification, and potential role in metabolism-related diseases.
Yirui LU ; Yutong RAN ; Hong LI ; Jiao WEN ; Xiaodong CUI ; Xiaoyun ZHANG ; Xiumei GUAN ; Min CHENG
Journal of Zhejiang University. Science. B 2023;24(12):1106-1122
With the development of modern sequencing techniques and bioinformatics, genomes that were once thought to be noncoding have been found to encode abundant functional micropeptides (miPs), a kind of small polypeptides. Although miPs are difficult to analyze and identify, a number of studies have begun to focus on them. More and more miPs have been revealed as essential for energy metabolism homeostasis, immune regulation, and tumor growth and development. Many reports have shown that miPs are especially essential for regulating glucose and lipid metabolism and regulating mitochondrial function. MiPs are also involved in the progression of related diseases. This paper reviews the sources and identification of miPs, as well as the functional significance of miPs for metabolism-related diseases, with the aim of revealing their potential clinical applications.
Humans
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Open Reading Frames
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Peptides
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Glucose
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Genome
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Metabolic Diseases
4.Molecular characterization of the feline T-cell receptor gamma alternate reading frame protein (TARP) ortholog.
Alexander Th A WEISS ; Marie Charlotte VON DEETZEN ; Werner HECHT ; Manfred REINACHER ; Achim D GRUBER
Journal of Veterinary Science 2012;13(4):345-353
T-cell receptor gamma alternate reading frame protein (TARP) is expressed by human prostate epithelial, prostate cancer, and mammary cancer cells, but is not found in normal mammary tissue. To date, this protein has only been described in humans. Additionally, no animal model has been established to investigate the potential merits of TARP as tumor marker or a target for adoptive tumor immunotherapy. In this study conducted to characterize feline T-cell receptor gamma sequences, constructs very similar to human TARP transcripts were obtained by RACE from the spleen and prostate gland of cats. Transcription of TARP in normal, hyperplastic, and neoplastic feline mammary tissues was evaluated by conventional RT-PCR. In felines similarly to the situation reported in humans, a C-region encoding two open reading frames is spliced to a J-region gene. In contrast to humans, the feline J-region gene was found to be a pseudogene containing a deletion within its recombination signal sequence. Our findings demonstrated that the feline TARP ortholog is transcribed in the prostate gland and mammary tumors but not normal mammary tissues as is the case with human TARP.
Animals
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Cats
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Continental Population Groups
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Humans
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Immunotherapy
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Models, Animal
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Open Reading Frames
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Prostate
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Prostatic Neoplasms
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Protein Sorting Signals
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Pseudogenes
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Reading Frames
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Receptors, Antigen, T-Cell
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Recombination, Genetic
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Spleen
5.Localization of functional domains of HEV ORF1 in cells.
Hui HUANG ; Zi-Zheng ZHENG ; Min ZHAO ; Jing-Xian LI ; Wang-Sheng LAI ; Ji MIAO ; Jun ZHANG ; Ning-Shao XIA
Chinese Journal of Virology 2011;27(3):195-201
To investigate the expression and localization of various functional domains of ORF1 polyprotein and ORF3 protein of hepatitis E virus in host cells, the coding sequences of the various functional domains (RdRp, HEL, MET, PLP, X) of ORF1 were separately cloned into pcDNA3. 1-GFP vectors for constructing the recombinant plasmids which were verified by enzyme digestion and sequencing. The exact expression of the fusion proteins were detected by Western Blot, and the distribution and localization were observed by the laser scanning confocal microscope(LSCM). In huh7 cells, GFP-RdRp proteins were found mainly in the nuclei, GFP-HEL proteins were distributed vesicularly around the nucleus, GFP-MET proteins were distributed granularly both in the nuclei and the cytoplasm, GFP-PLP proteins had polar distribution around the nucleus, and unknown GFP-X proteins were distributed uniformly both in the nuclei and the cytoplasm. Different localization of these proteins verified the previous data obtained from in vitro studies, providing a support for further research on the biological functions of various proteins coded by HEV genome.
Blotting, Western
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Cells, Cultured
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Hepatitis E virus
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genetics
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Humans
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Open Reading Frames
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Viral Proteins
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genetics
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physiology
6.ORF Miner: a Web-based ORF Search Tool.
Genomics & Informatics 2009;7(4):217-219
The primary clue for locating protein-coding regions is the open reading frame and the determination of ORFs (Open Reading Frames) is the first step toward the gene prediction, especially for prokaryotes. In this respect, we have developed a web-based ORF search tool called ORF Miner. The ORF Miner is a graphical analysis utility which determines all possible open reading frames of a selectable minimum size in an input sequence. This tool identifies all open reading frames using alternative genetic codes as well as the standard one and reports a list of ORFs with corresponding deduced amino acid sequences. The ORF Miner can be employed for sequence annotation and give a crucial clue to determination of actual protein-coding regions.
Amino Acid Sequence
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Animals
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Ecthyma, Contagious
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Genetic Code
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Open Reading Frames
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Resin Cements
7.Characterization of a Potential Pathogenicity Island of Enterotoxigenic Bacteroides fragilis (ETBF) Strains.
Journal of the Korean Society for Microbiology 1998;33(3):273-280
Enterotoxigenic B. pagilis (ETBF) strains which produce a 20 kDa zinc metalloprotease toxin (BFI) have been associated with diarrheal disease of animals and young children. Using B. pngilis toxin gene (bfi) from strain 86-5443-2-2 (piglet isolate) as a probe, the gene was identified in 74/77 human and animal ETBF strains but only 2/97 non-toxigenic B. fragilis (NTBF) strains. The region flanking bp was mapped with several restriction enzymes and 8 resriction fragments aacent to bft were used to probe colony blots of 77 KTBF and 97 NTBF strains. All 74 bft-positive ETBF strains hybridized to the 8 probes spanning a ca. 18 kb chromosomal region; however, this 18 kb region was absent in the 3 ETBF strains lacking p, and 47 of the 97 (48%) NTBF strains lacked the entire 18 kb region. Of note, the 2 NTBF strains containing btf did not have a ca. 12 kb region upstream of btfp. A ca. 9 kb fragment flanking the btf gene has been sequenced. Analysis of this data revealed several open reading frames (ORF) of which 3 are of particular interest (ORFs 1, 2 and 3). ORF1 and ORF3 encode proteins with significant homology to mobilization proteins, and ORF2 encodes a protein with significant homology to metalloprotease proteins, but only 50% similarity and 30% identity to BFf. These results suggest: 1) the btf genes are flanked by at least 18 kb of DNA largely unique to ETBF strains indicating a putative pathogenic island, 2) another metalloprotease protein present in ETBF strains may contribute to the pathogenicity and variable virulence of these diarrheagenic strains and 3) the pathogenic island may be mobiTized among different Bacteroides strains, and possibly among different species of intestinal bacteria.
Animals
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Bacteria
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Bacteroides fragilis*
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Bacteroides*
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Child
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DNA
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Genomic Islands*
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Humans
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Open Reading Frames
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Virulence*
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Zinc
8.HExDB: Human EXon DataBase for Alternative Splicing Pattern Analysis.
Junghwan PARK ; Minho LEE ; Jong BHAK
Genomics & Informatics 2005;3(3):80-85
HExDB is a database for analyzing exon and splicing pattern information in Homo sapiens. HExDB is useful for specific purposes: 1) to design primers for exon amplification from cDNA and 2) to understand the change of ORFs by alternative splicing. HExDB was constructed by integrating data from AltExtron which is the computationally predicted exon database, Ensemble cDNA annotation, and Affymetrix genome tile published recently. Although it may contain false positive data, HExDB is good starting point due to its sensitivity. At present, there are as many as 2,046,519 exons stored in the HExDB. We found that 16.8% of the exons in the database was constitutive exons and 83.1% were novel gene exons.
Alternative Splicing*
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Animals
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DNA, Complementary
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Ecthyma, Contagious
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Exons*
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Genome
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Humans*
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Open Reading Frames
9.Protein Expression of the Human Norovirus Capsid Gene using the Baculovirus Expression System.
Ji Young JIN ; Chul Jong PARK ; Seung Won PARK ; Soon Young PAIK
Journal of Bacteriology and Virology 2011;41(3):183-187
Human norovirus (HuNoV) is the major etiological agent of nonbacterial gastroenteritis worldwide. However, due to the absence of a rapid and sensitive diagnostic system, detection and monitoring have been limited. The HuNoV genome is composed of three open reading frames (ORFs). And major capsid protein, ORF2, is designated as a viral protein 1 (VP1). In this study, the baculovirus expression system was used for expression of the HuNoV capsid protein, VP1. Recombinant baculoviruses can be used as potent tools in HuNoV studies.
Baculoviridae
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Capsid
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Capsid Proteins
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Gastroenteritis
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Genome
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Humans
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Norovirus
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Open Reading Frames
10.Morphological Transformation Region II (mtrII) of Human Cytomegalovirus.
Yeon Myung SHIN ; Soo Sang SOHN ; Joong Shin KANG ; Sung Ik CHANG ; In Hwan LEE
Journal of the Korean Cancer Association 1997;29(3):495-503
PURPOSE: Human herpesviruses have been associated with the etiology of several human cancers. The role of these viruses in carcinogenesis has not yet been clarified. This study focused on identifying and characterizing the transforming potential of cloned DNA fragments from human cytomegalovirus (HCMV). MATERIALS AND METHODS: Multiple DNA fragments of HCMV were applied to cells for transformation. Morphological transforming region II (mtrII) of HCMV strain Towne has been identified to a 3.0kb XbaI-BamHI DNA fragment which was retained in transformed cells. The transforming activity was induced by a 980 bp BaII-Xho I subfragment (pBS980) containing both promoter/ regulatory elements as well as three open reading frames (ORFs), i.e., 79ORF, 83ORF, and 34ORF. The ORFs have been evaluated for transforming potential in NIH3T3 cells. RESULTS: MtrII (pBS980) has BglII restriction enzyme site which divides into two subfragments, pBS440 and pBS540, the latter has whole 83ORF, 34ORF, and fragment of 79ORF, the former has only fragment of 79ORF. Among three ORFs, 83ORF and 34ORF were not functional in transformation, because in pBS540 these ORFs were not truncated. CONCLUSION: The 79ORF (79-aa transforming peptide) has allowed a better approach to determine the role of HCMV in human carcinogenesis.
Animals
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Carcinogenesis
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Clone Cells
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Cytomegalovirus*
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DNA
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Ecthyma, Contagious
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Herpesviridae
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Humans*
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Open Reading Frames