• Title/Summary/Keyword: N gene

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ON THE REPRESENTATION OF PROBABILITY VECTOR WITH SPECIAL DIFFUSION OPERATOR USING THE MUTATION AND GENE CONVERSION RATE

  • Choi, Won
    • Korean Journal of Mathematics
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    • v.27 no.1
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    • pp.1-8
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    • 2019
  • We will deal with an n locus model in which mutation and gene conversion are taken into consideration. Also random partitions of the number n determined by chromosomes with n loci should be investigated. The diffusion process describes the time evolution of distributions of the random partitions. In this paper, we find the probability of distribution of the diffusion process with special diffusion operator $L_1$ and we show that the average probability of genes at different loci on one chromosome can be described by the rate of gene frequency of mutation and gene conversion.

The DNA region of rtn gene essential for resistance against N4 infection (N4에 대해 내성을 나타내는데 필요한 rtn 유전자의 부위)

  • 이동환;유선미;황의욱;이영훈;채건상
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.290-295
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    • 1991
  • N4 phage, which infects E. coli K-12 strains, could not infect E. coli K-12 strains containing rtn(resistant to N4) gene on plasmids, which was isolated from Proteus vulgaris ATCC 13315. The region of rtn gene for Rtn phenotype was reduced to the 1.7 kb HincII-AccI fragment, and rtn gene seemed to have its own promoter. This putative promoter was present in 107 bp HindII-DraI fragment, and known to be functional in E. cole K-12, which is supported by the fact that phenotype of a subclone, pRMG103A1B which does not contain the 107 bp fragment, was dependent on the existance of a functional promoter in the upstream of rtn gene, and that the 107 bp fragment had promoter activity when located in the upstream of structural gene of galactodinase of E. coli. The promoter-bearing fragment contains two overlapping putative promoter sequences, both of which show a fit in eight of twelve nucleotides with consensus sequences of E. coli promoters at the -35 and -10 regions.

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Expression of Neurotensin/Neuromedin N Precursor in Murine Mast Cells

  • Ahn, Hyun-Jong;Cho, Jeong-Je
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.6
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    • pp.495-501
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    • 2001
  • We have cloned the mouse neurotensin/neuromedin N (NT/N) gene from the murine mast cell line Cl.MC/C57.1 for the first time. The murine NT/N cDNA clone consisted of 765 nucleotides and coded for 169 peptide residues with an N-terminal signal peptide, and the C-terminal region contained of one copy of neurotensin (NT) and one copy of neuromedin N (NN). Total of four Lys-Arg dibasic motifs were present; one each at the middle of the open reading frame, at the N-terminal of NN, at the C-terminal of NT, and between NN and NT. Amino acid sequence analysis of the mouse NT/N revealed 90% homology to that of the rat NT/N gene. NT/N is expressed in murine mast cell lines (Cl.MC/C57.1 and P815), but not in murine bone marrow-derived mast cells (BMMCs), murine macrophage cell line (RAW 264.7), nor in murine T cell line (EL-4). NT/N mRNA in C1.MC/C57.1 is highly inducible by IgE cross-linking, phorbol myristate acetate, neurotensin, and substance P. Following the treatment of demethylating agent, 5-azacytidine (5-azaC), the NT/N gene was induced in BMMCs in response to IgE cross-linking. 5-azaC-treated BMMCs did not express the NT/N gene without additional stimuli. These findings suggested that the regulation of NT/N gene expression was dependent on the effects of not only gene methylation but also enhancer and/or repressor proteins acting on the NT/N promoter.

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Novel polymorphisms of dopa decarboxylase gene and their association with lamb quality traits in Indonesian sheep

  • Ratna Sholatia Harahap;Ronny Rachman Noor;Yuni Cahya Endrawati;Huda Shalahudin Darusman;Asep Gunawan
    • Animal Bioscience
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    • v.36 no.6
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    • pp.840-850
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    • 2023
  • Objective: This study aimed to investigate the polymorphisms of the dopa decarboxylase (DDC) gene and association analysis with lamb quality and expression quantification of the DDC gene in phenotypically divergent Indonesian sheep. Methods: The totals of 189 rams with an average body weight of 24.12 kg at 10 to 12 months were used to identify DDC gene polymorphism using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Among 189 rams, several rams representing various sheep genotypes were used for an association study between genotypes and phenotypic traits with proc general linear model (GLM) analysis. In addition, the gene expression analysis of the DDC mRNA in the phenotypically divergent sheep population was analyzed using quantitative reverse-transcription PCR. Results: The DDC gene (g. 5377439 G>A) showed polymorphisms that indicated three genotypes: AA, AG, and GG. The DDC gene polymorphism was significantly associated (p≤0.05) with carcass characteristics including carcass percentage, carcass length, hot and cold carcass; physical properties of lamb quality including pH value; retail cut carcass; fatty acid composition such as fat content, pentadecanoic acid (C15:0), tricosylic acid (C23:0), lignoceric acid (C24:0), oleic acid (C18:1n9c), elaidic acid (C18:1n9t), nervonic acid (C24:1), linoleic acid (C18:2n6c), arachidonic acid (C20:4n6), cervonic acid (C22:6n3); and mineral content including potassium (K). The GG genotype of the DDC gene had the best association with lamb quality traits. The DDC gene expression analysis mRNA showed no significant difference (p≥0.05) between lamb quality traits. Conclusion: The DDC gene could be used as a potential candidate gene to improve lamb quality.

Application of the melanocortin 1 receptor (MC1R) gene for discrimination of Hanwoo from Holstein beef using real-time polymerase chain reaction (PCR)

  • Ra, Do-Kyung;Lee, Sung-Mo;Park, Eun-Jeong;Lee, Jung-Goo
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.557-562
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    • 2007
  • This study was carried out to discriminate Hanwoo from the milking and hybrid cattle by detection of MC1R gene related to bovine hair color. One hundred sixty six samples were collected from the abattoir (n = 106) and local market (n = 60). The beef from abattoir were originated from Hanwoo (n=27), Holstein (n=29), Hybrid (n=45) and imported cattle (n=5), respectively. The beef from market consisted of Hanwoo (n=36), Holstein (n=7) and imported ones (n=17). Commercialized screening kit (Kogenebiotec, Korea) was used for MC1R gene analysis. As a result, Hanwoo was discriminated from Holstein. However, 9 of 45 hybrid and 11 of 22 imported beef samples were indistinguishable from Hanwoo. It could be explained by second generation of crossing of Hanwoo with Holstein or the cattle with silver or yellow hair. This results suggest that additional tests as well as MC1R gene detection be needed to confirm Hanwoo beef among cattle beef.

Characterization of a Chitinase Gene Exhibiting Antifungal Activity from a Biocontrol Bacterium Bacillus licheniformis N1

  • Lee, Kwang-Youll;Heo, Kwang-Ryool;Choi, Ki-Hyuck;Kong, Hyun-Gi;Nam, Jae-Sung;Yi, Young-Byung;Park, Seung-Hwan;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.344-351
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    • 2009
  • A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

Expression of the FLP recombinase of the 2 $\mu$m plasmid of yeast in the cultured cells of Bombyx mori using a transient expression vector (Yeast 2 $\mu$m 플라스미드 유래 FLP recombinase 유전자의 곤충 배양세포내 발현)

  • 강석우;윤은영
    • Journal of Sericultural and Entomological Science
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    • v.39 no.1
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    • pp.36-43
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    • 1997
  • In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

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Detection of invA and spvC in Salmonella spp. isolated from duck farms (오리 농장에서 분리한 Salmonella속 균에서 invA 및 spvC gene의 검출)

  • Cho, Jae-Keun
    • Korean Journal of Veterinary Service
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    • v.33 no.4
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    • pp.341-344
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    • 2010
  • Poultry and poultry products have been implicated as a major source of Salmonella infection in human, and infection due to Salmonella serotypes continue to be a major health problem. The presence of two virulence genes, invA and spvC, in 34 Salmonella isolates obtained from duck farms was investigated. All isolates contained the invA gene, and spvC gene was found in 20 (58.8%) of 34 Salmonella isolates : S. Typhimurium (n=8), S. Fyris (n=5), S. Enteritidis (n=3), S. Typhimurium var. copenhagen (n=1), S. Haardt (n=1) and S. Mbandaka (n=1). This study showed the presence of the spvC gene was widely distributed in between different Salmonella enterica isolates.

Molecular and Biochemical Studies on the DNA Replication of Bacteriophage T7: Functional Analysis of Amino-terminal Region of Gene 2.5 Protein

  • Kim, Young-Tae;Lee, Sung-Gu;Kim, Hak-Jun
    • BMB Reports
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    • v.28 no.6
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    • pp.484-489
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    • 1995
  • The product of bacteriophage T7 gene 2.5 is a single-stranded DNA binding protein and plays an important role in T7 DNA replication, recombination, and repair. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth (Kim and Richardson, 1993). The C-terminal truncated gene 2.5 protein ($GP2.5-{\Delta}21C$) cannot substitute for wild-type gene 2.5 protein in vivo; suggesting that the C-terminal domain of gene 2.5 protein is essential for protein-protein interactions (Kim and Richardson, 1994; J. Biol. Chem. 269, 5070-5078). Truncated gene 2.5 proteins lacking 19 residues ($GP2.5-{\Delta}19N$) and 39 residues ($GP2.5-{\Delta}39N$) from the amino-terminal domain were constructed by in vitro mutagenesis. $GP2.5-{\Delta}19N$ can support the growth of T7 phage lacking gene 2.5 while $GP2.5-{\Delta}39N$ cannot substitute for wild-type gene 2.5 protein in vivo; however, its ability to bind to single-stranded DNA is not affected. These results clearly demonstrate that the 20~39 amino-terminal region of gene 2.5 protein is required for T7 growth in vivo but may not be involved in DNA binding activity.

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Improving siRNA design targeting nucleoprotein gene as antiviral against the Indonesian H5N1 virus

  • Hartawan, Risza;Pujianto, Dwi Ari;Dharmayanti, Ni Luh Putu Indi;Soebandrio, Amin
    • Journal of Veterinary Science
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    • v.23 no.2
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    • pp.24.1-24.10
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    • 2022
  • Background: Small interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia. Objectives: The main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus. Methods: The effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells. Results: The siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene. Conclusions: These findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.