• Title/Summary/Keyword: nucleotide sequence homology

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Cloning and Sequence Analysis of the trpB, trpA and 3' trpC(F) Gens of Vibrio metschnikovii Strain RH530 (Vibrio metschnikovii 균주 RH530의 trpB, trpA 그리고 3' trpC(F) 유전자의 클로닝 및 염기서열 결정)

  • Kwon, Yong-Tae;Kim, Jin-Oh;Yoo, Young-Dong;Rho, Hyune-Mo
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.120-125
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    • 1994
  • The genes, trpB, trpA and 3’ trpC(F) of Vibrio metschnikovii strain RH530 were cloned and sequenced. The trpB and trpA genes had open reading frames of 1,173 bp and 804 bp encoding 391 and 268 amino acids, respectively. The trpB and trpA genes had conventional ribosome-binding sequences and overlapped with each other by one nucleotide, suggesting that these two genes are translationally coupled. 115 nucleotide upstream the trpB start codon, tjere was an incomplete open reading frame of the 3’-end of the trpC(F). The amino acid sequences of trpB, trpA and trpC(F) of V. metschnikovii RH530 had identities of 64.2%, 82.4% and 73.7% respectively, for those of V. parahaemolyticus; 58.7%, 72.3% and 54.9%, respectively, for Salmonella typhimurium; and 42.6%. 54.1% and 12.5%, respectively, for brevibacterium lactofermentum. The genetic organization of these genes, especially in the noncoding region between trpC(F) and trpB, was distinct from that of Enterobacteriaceae.

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Identification of the sprU Gene Encoding an Additional sprT Homologous Trypsin-Type Protease in Streptomyces griseus

  • YANG HYE-YOUNG;CHOI SI-SUN;CHI WON-JAE;KIM JONG-HEE;KANG DAE-KYUNG;CHUN JAESUN;KANG SANG-SOON;HONG SOON-KWANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1125-1129
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    • 2005
  • Cloning of a 6.6-kb BamHI digested chromosomal DNA from S. griseus IFO13350 revealed the presence of an additional gene encoding a novel trypsin-like enzyme, named SprU. The SprU protein shows a high homology ($79\%$ identity, $88\%$ similarity) with the SGT protease, which has been reported as a bacterial trypsin in the same strain. The amino acid sequence deduced from the nucleotide sequence of the sprU gene suggests that SprU is produced as a precursor consisting of an amino-terminal presequence (29 amino acid residues), prosequence (4 residues), and mature trypsin consisting of 222 amino acids with a molecular weight of 22.94 kDa and a calculated pI of 4.13. The serine, histidine, and aspartic acid residues composing the catalytic triad of typical serine proteases are also well conserved. When the trypsin activity of the SprU was spectrophotometrically measured by the enzymatic hydrolysis of the artificial chromogenic substrate, N-${alpha}$-benzoyl-DL-arginine-p-nitroanilide, the S. lividans transformant with pWHM3-U gave 3 times higher activity than that of control. When the same recombinant plasmid was introduced into S. griseus, however, the gene dosage effect was not so significant, as in the cases of other genes encoding serine proteases, such as sprA, sprB, and sprD. Although two trypsins, SprU and SGT, have a high degree of homology, the pI values, the gene dosage effect in S. griseus, and the gene arrangement adjacent to the two genes are very different, suggesting that the biochemical and biological function of the SprU might be quite different from that of the SGT.

Cloning, Sequencing, and Expression of the Gene Encoding a Multidomain Endo-$\beta$-1,4-Xylanase from Paenibacillus curdlanolyticus B-6, and Characterization of the Recombinant Enzyme

  • Waeonukul, Rattiya;Pason, Patthra;Kyu, Khin Lay;Sakka, Kazuo;Kosug, Akihiko;Mori, Yutaka;Ratanakhanokchai, Khanok
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.277-285
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    • 2009
  • The nucleotide sequence of the Paenibacillus curdlanolyticus B-6 xyn10A gene, encoding a xylanase Xyn10A, consists of 3,828 nucleotides encoding a protein of 1,276 amino acids with a predicted molecular mass of 142,726 Da. Sequence analysis indicated that Xyn10A is a multidomain enzyme comprising nine domains in the following order: three family 22 carbohydrate-binding modules (CBMs), a family 10 catalytic domain of glycosyl hydrolases (xylanase), a family 9 CBM, a glycine-rich region, and three surface layer homology (SLH) domains. Xyn10A was purified from a recombinant Escherichia coli by a single step of affinity purification on cellulose. It could effectively hydrolyze agricultural wastes and pure insoluble xylans, especially low substituted insoluble xylan. The hydrolysis products were a series of short-chain xylooligosaccharides, indicating that the purified enzyme was an endo-$\beta$-1,4-xylanase. Xyn10A bound to various insoluble polysaccharides including Avicel, $\alpha$-cellulose, insoluble birchwood and oat spelt xylans, chitin, and starches, and the cell wall fragments of P. curdlanolyticus B-6, indicating that both the CBM and the SLH domains are fully functioning in the Xyn10A. Removal of the CBMs from Xyn10A strongly reduced the ability of plant cell wall hydrolysis. These results suggested that the CBMs of Xyn10A play an important role in the hydrolysis of plant cell walls.

Molecular Cloning and Nucleotide Sequence of Connexin 35 cDNA in the Ovary from the Sweetfish, Plecoglossus altivelis (은어, Plecoglossus altivelis 난소에서 발현하는 Connexin 35 cDNA의 해석)

  • Choi, Cheol-Young;Chang, Young-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.565-571
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    • 2000
  • Mixed primers based on the high sequence homology of selected regions of known connexins (Cxs) was used for PCR reaction. A full-length connexin cDNA of sweetfish (Plecoglossus altivelis) was cloned by rapid amplification of cDNA 5'and (5'RACE) and 3'RACE method. When compared to other known Cx sequences, homology of sweetfish Cx cDNA to Atlantic croaker, Mycropogonias undulatus Cx32.7, bovine, Bos taurus Cx44 and Atlantic croaker Cx32.2 were $63.8{\%},\;61.6{\%}\;and\;56.7{\%}$, respectively. This cDNA encoded 308 amino acids (35,028 dalton) and named as sweetfish Cx35. Hydropathicity analysis of predicted amino acid sequences indicated that sweetfish Cx35 have four major hydrophobic regions and four major hydrophilic regions, suggesting its topology is similar to that of known Cxs. The presence of a tfical Cx consensus sequences were identified in each of the extracellular loops (first loop and second loop).

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Brain invasion of bovine coronavirus: molecular analysis of bovine coronavirus infection in calves with severe pneumonia and neurological signs

  • Semaha Gul Yilmaz;Ozge Aydin;Hasan Emre Tali;Gizem Karadag;Kivilcim Sonmez;Erhan Bayraktar;Aysun Yilmaz;Nuri Turan;Zihni Mutlu;Munir Iqbal;Jurgen A. Richt;Huseyin Yilmaz
    • Journal of Veterinary Science
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    • v.25 no.4
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    • pp.45.1-45.12
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    • 2024
  • Importance: Although the role of bovine coronavirus (BCoV) in calf diarrhea and respiratory disorders is well documented, its contribution to neurological diseases is unclear. Objective: This study conducted virological investigations of calves showing diarrhea and respiratory and neurological signs. Methods: An outbreak of diarrhea, respiratory, and neurological disorders occurred among the 12 calves in July 2022 in Istanbul, Türkiye. Two of these calves exhibited neurological signs and died a few days after the appearance of symptoms. One of these calves was necropsied and analyzed using molecular and histopathological tests. Results: BCoV RNA was detected in the brain, lung, spleen, liver, and intestine of the calf that had neurological signs by real-time reverse transcription polymerase chain reaction. Immunostaining was also observed in the intestine and brain. A 622 bp S1 gene product was noted on gel electrophoresis only in the brain. Phylogenetic analysis indicated that the BCoV detected in this study had a high proximity to the BCoV strain GIb with 99.19% nucleotide sequence homology to the strains detected in Poland, Israel, Türkiye, and France. No distinct genetic lineages were observed when the brain isolate was compared with the respiratory and enteric strains reported to GenBank. In addition, the highest identity (98,72%) was obtained with the HECV 4408 and L07748 strains of human coronaviruses. Conclusions and Relevance: The strain detected in a calf brain belongs to the GIb-European lineage and shares high sequence homology with BCoV strains detected in Europe and Israel. In addition, the similarity between the human coronaviruses (4408 and L07748) raises questions about the zoonotic potential of the strains detected in this study.

First Report on Carnation vein mottle virus in Dianthus barbatus in Korea

  • Chung, B.N.;Kim, B.D.;Choi, G.S.;Kim, J.S.
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.224-228
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    • 2004
  • A potyvirus causing chlorotic mottle and yellow spots on leaves of Dianthus barbatus was isolated and identified as an isolate of Carnation vein mottle virus (CVMV). Purified preparations of Chenopodium quinoa infected with CVMV-K showed filamentous particles between 695 and 785 om long. Many cytoplasmic inclusions were observed, and these consisted of pinwheels, dense bands, loops, and circles. The coat protein of CVMV-K was about 32 KDa in western blot analysis using a CVMV antibody. The nucleotide sequence of coat protein gene showed 97.6% homology with a Japanese isolate. The genome size of CVMV-K was about 9.0 kb by dsRNA analysis. These results indicate that the virus is an isolate of CVMV. This is the first report on CVMV in Korea.

First Report on Poinsettia mosaic virus in Korea

  • Chung, B.N.;Lee, E.K.;Jeong, M.I.;Kim, H.R.
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.220-223
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    • 2004
  • Most plants of commercial poinsettia cultivars grown from cuttings develop mosaic and chlorotic dot symptoms on leaves. Reverse transcription-polymerase chain reaction (RT-PCR) test showed that they were infected with Poinsettia mosaic virus (PnMV). In a survey of commercially grown poinsettias conducted in Korea, PnMV was detected in ten of ten poinsettia cultivars sampled and in 100% of 178 samples tested. The virus has isometric particles and about 29 nm in diameter. Crystalline virus particles were observed in cytoplasm of cells of diseased plants by transmission electron microscopy. Nucleotide sequence of coat protein gene of PnMV- Kl showed 97.3% homology with that of a German isolate. This is the first report on PnMV in Korea.

Physical Analysis of nahQ tnpA Genes from Pseudomonas fluorescens

  • Seol, Ja-Young;Chol, Soon-Young;Min, Kyung-Hee
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.338-342
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    • 2001
  • Pseudomonas fluorescens SM11 is a naphthalene-degrading strain whose dissimilatory genes are cho-mosomally encoded. We have cloned the 2.9 kb Sal I fragment harboring genes for the naphthalene-degrading upper pathway. The nucleotide sequences were determined to be nahQ, napA, and partial regions of nahE genes. The nahQ encods a protein of 188 amino acid residues with a deduced molec-ular wight of 20.8kDa. The high homology with other proteins suggests that NAhQ may be an active and useful protein whtich gives as selective advantage to naphthalene degradatin. Transposase(TnpA)encodes a polypetide chain with a molecular mass of 41.8kDa consisting of 376 amino acid residues. The deduced anino acid sequence of tnpA revealed 96% idenitity with putative transposase of P. stutzeri OX1,. It was assumed that transposase plays an important role in the evloution of the catabloic-path way in the regulation of nah expression.

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Isolation and Characterization of the sod2$^{2+}$ Gene Encoding a Putative Mitochondrial Manganese Superoxide Dismutase in Schizosaccharomyces bombe

  • Jeong, Jae-Hoon;Kwon, Eun-Soo;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.37-41
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    • 2001
  • The fission yeast Schizosaccharomyces pombe contains two distinct superoxide dismutase (SOD) activities, one in the cytosol encoded by the $sod2^{+}$ gene and the other in mitochondria. The $sod2^{+}$ gene encoding putative mitochondrial manganese superoxide dismutase (MnSOD) was isolated from the S. pombe genomic library using a PCR fragment as the probe. The nucleotide sequence of the $sod2^{+}$ gene and its flanking region (4051 bp HindIII fragment) was determined. An intron of 123 nt in size was predicted and confirmed by sequencing the cDNA following reverse transcription PCR. The predicted Sod2p consists of 218 amino acid residues with a molecular mass of 24,346 Da. The deduced amino acid sequence showed a high degree of homology with other MnSODs, especially in the metal binding residues at the active site and their relative positions. The transcriptional start site was mapped by primer extension at 231 at upstream from the ATG codon. A putative TATA box(TATAAAA) was located 58 nt upstream from the transcriptional start site and putative polyadenylation sites were located at 1000, 1062, and 1074 nt downstream from the ATG start codon.

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Sequence analysis of segment A gene of a very virulent infectious bursal disease virus recently isolated in Korea (최근 국내 분리 고병원성 infectious bursal disease virus의 segment A 유전자 특성)

  • Oh, Hyun Seok;Lee, Jin Hwa;Kwon, Hyuk Moo;Sung, Haan Woo
    • Korean Journal of Veterinary Research
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    • v.51 no.1
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    • pp.37-46
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    • 2011
  • Infectious bursal disease virus (IBDV) is a member of the Avibirnavirus genus of the Birnaviridae family which genome consists of two segments (A and B) of double stranded RNA. Segment A gene of KNU08010 isolate, which was isolated from a 15-day-old chicken flock in 2008, was sequenced and compared with other IBDV isolates including SH/92 strain, the first Korean very virulent (vv) IBDV isolate. The amino acid sequences of segment A gene showed that KNU08010 had 99.2% homology with SH92 strain. KNU08010 isolate had specific amino acids A222, I242, I256, I294 and S299 which are highly conserved among vvIBDV strains. Phylogenetic analysis based on the nucleotide sequences of variable region of the VP2 gene of 18 IBDV strains revealed that KNU08010 was grouped with vvIBDVs and was closely related to Korean vvIBDVs isolated from wild birds.