• Title/Summary/Keyword: M/BP/1

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Cloning and Nucleotide Sequence Analysis of the M Protein of a Korean Isolate of Infectious Hematopoietic Necrosis Virus (한국에서 분리된 전염성 조혈괴저바이러스의 M 단백질의 유전자 클로닝과 염기서열 분석)

  • Park, Jeong-Min;Kim, Hyun-Ju;Mun, Chang-Hoon;Cho, Wha-Ja;Cha, Seung-Ju;Yoon, Won-Joon;Park, Jeoug-Jae;Lee, Eun-Hee;Park, Myoung-Ae;Sohn, Sang-Gyu;Park, Jeong-Woo
    • Korean Journal of Microbiology
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    • v.34 no.1_2
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    • pp.64-68
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    • 1998
  • In order to identify the characteristics of a Korean isol ate of infectious hematopoietic necrosis virus(IHNV), IHNV-PRT, we have cloned and analyzed cDNAs coding for matrix protein M1 and M2 of the IHNV-PRT. The Ml gene contained 693 bp open reading frame and encoded a protein of 230 amino acids with a molecular weight of 25.9 kDa. The M2 gene had 588 bp open reading frame, encoding a protein of 195 amino acids with a molecular weight of 21.9 kDa. On the deduced amino-acid sequences, M1 and M2 of the IHNV-PRT were found to be 92-93% (M1) and 97% (M2) identical to those of foreign isolates of IHNV. These results indicate that M genes of the IHNV are highly conserved among different strains of IHNV.

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Characterization of the Mutant of Streptomyces sp. SL-387(KCTC 0102BP) Producing Aminopeptidase M Inhibitors (Aminopeptidase M 저해제를 생산하는 Streptomyces sp. SL-387 (KCTC 0102BP) 변이주의 특성)

  • Chung, Myung-Chul;Chun, Hyo-Kon;Lee, Ho-Jae;Lee, Choong-Hwan;Kho, Yung-Hee
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.47-52
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    • 1995
  • Since the original productivity of new aminopeptidase M inhibitors MR-387A and B by Streptomyces sp. SL-387 (KCTC 0102BP) was not enough for further chemical and biological evaluation, mutation of parent strain by the treatment of N-methyl-N'-nitro-N-nitrosoguanidine was performed in order to obtain a clone with greater inhibitory activity. Mutant N-3 was selected due to a 6-fold greater productivity (40 $\mu$g/ml) than that of the wild type(6.7 $\mu$g/ml). This mutant was resistant to 3,4-dehydro-DL-proline, an antimetabolite of proline, with 25 $\mu$g/ml of minimum inhibitory concentration. Furthermore, the characteristic morphological change from spiral spore chain in wild type to straight in mutant was observed. An aminopeptidase M nhibitor different from MR-387A and B was isolated from the culture broth of the mutant. This inhibitor was composed of 2 proline, 1 valine, and an unknown amino acid which is presumably 3-amino-4-phenylbutanoic acid. IC$_{50}$ value (89.1 $\MU$g/ml) of the purified inhibitor was lower than that of other inhibitors, which may be due to the absence of 2(S)-hydroxyl group within the structure of 3-amino-4-phenyl- butanoic acid.

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Clonorchis sinensis tropomyosin: Cloning and sequence of partial cDNA amplified by PCR (간흡충 tropomyosin: PCR로 일부분 증폭된 cDNA의 cloning 및 염기서열)

  • 홍성종
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.285-292
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    • 1993
  • C. sinensis total RMh was containing large amount of 185 rRNA but little 285 rRNA. The size of the double-stranded cDNA synthesized from poly $(A)^{+}$ mRNA was 0.4-4.2 kb long with tapering unto 9.5 kb. Degenerated oligonucleotides (as 2 sense and 3 antisense Primers) were designed on the conserved regions of the known tropomyosin amino acid sequences. From one out of the PCR amplifications using total CDNA and matrix of primers, a specific gene product, 580 bp in size, was produced. Upon Southern hybridization of the PCR products with Schistosomn mnnsoni tropomyosin (SMTM) CDNA, only one signal appeared at the band of 580 bp product. This 580 bp product was considered to encode C. sinensis tropomyosin (CSTM) and cloned in pGEM-3Zf(-) for DNA sequencing. CSTM cDNA was 575 bp containing one open reading frame of 191 predicted amino acids, which revealed 86.3% homology with SMTM and 51.1% with rrichostronsylur coeubnlormis tropomyosin. CSTM cDNA obtained will serve as a probe in the studies of molecular cloning of CSTM.

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HspBP1 Is the Negative Regulator of the Bovine Progesterone Receptor

  • Park, K.M.;Song, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.9
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    • pp.1261-1267
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    • 2003
  • We have investigated whether HspBP1, a Hsp70 binding protein, could have effect on the assembly of the bovine progesterone receptor (bPR) with a chaperone complex consisting of bovine Hsp90 (bHsp90), bovine Hsp70 (bHsp70), Hop, Ydj-1, and p23. The bPR, isolated in its native conformation, loses its function to interact with progesterone hormone in the absence of this protein complex. However, in the presence of bHsp90, bHsp70, Hop, p23 and Ydj-1, its function could be restored in vitro. Our findings here indicate that the inclusion of HspBP1 to five-protein system prevented the proper assembly of progesterone receptor-chaperone complex and induce the loss of bPR ability to interact with hormone. Immunoprecipitation assays of bPR with HspBP1 show that the presence of HspBP1 did not have any effect on the assembly of Ydj-1 and bHsp70 with the progesterone receptor. However, further assembly of Hsp90, Hop and p23 was completely prevented and the function of the bPR was lost. In vitro competition and protein folding assays indicated that the binding of HspBP1 to bHsp70 prevented the ternary complex formation of bHsp70, bHsp90, and Hop. These results indicate that HspBP1 is a negative regulator of the assembly of Hsp90, Hop and Hsp70, and thus, prevent the proper maturation of unliganded bPR with chaperones assembly system.

Determination of Phthalate Metabolites in Korean Children's Urine by High Performance Liquid Chromatography with Triple Quadrupole Tandem Mass Spectrometry (LC/MS/MS를 이용한 어린이 소변 중 프탈레이트 대사산물 분석)

  • Kho, Young-Lim;Jeong, Jee-Yeon;Choi, Kyung-Ho;Kim, Pan-Gyi
    • Journal of Environmental Health Sciences
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    • v.34 no.4
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    • pp.271-278
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    • 2008
  • Dialkylated phthalates have been commonly used as plasticizers and a variety of applications. Phthalate diesters have been shown to be developmental and reproductive toxicants. It is very difficult to exactly estimate the dose of dialkylated phthalates taken up by the general population because of environmental contamination. Urinary metabolites of phthalates enabled to estimate internal exposure. The objective of this study was quantitative determination of phthalate metabolites by LC/MS/MS with on-line cleanup method to analyze phthalate metabolites in Korean children's urine. We employed LC/MS/MS with on-line enrichment and column-switching techniques for this biological monitoring. Metabolites determined were 4 primary metabolites; MEHP, MnBP, MiBP, MEP and 2 secondary metabolites of DEHP; 5-OH-MEHP), 5-oxo-MEHP. We analyzed children's urine from 30 boys and 30 girls. The method detection limit of phthalate metabolites were 0.03 ng/mL for MEP, 1.05 ng/mL for MBP, 0.22 ng/mL for MEHP, 0.15 ng/mL for 5-OHMEHP and 0.16 ng/mL for 5-oxo-MEHP, respectively. Switching Column LC/MS/MS was proven to be a useful tool to determine metabolites of phthalate diesters in human urine. The correlation among phthalate metabolites was very high and statistically significant, except MEP. The children's age (months) was negatively correlated to the concentration of phthalate metabolites. The geometric mean concentration of phthalate metabolites (mg/g creatinine) in children's urine were 25.5 for MEP, 130.3 for MnBP, 56.8 for MiBP, 19.5 for MEHP, 85.6 for 5-OH-MEHP and 83.1 for 5-oxo-MEHP, respectively. Levels of estimated daily intake of parent phthalate compounds (${\mu}g$/kg bw/day) were 0.8 for DEP, 5.0 for DnBP, 1.9 for DiBP and $8.9{\sim}14.2$ for DEHP, respectively. Estimated daily intake for DEP and DiBP were lower than those of other studies but the value for DEHP was higher than that of other study.

Paralytic Peptide Binding Protein (PP-BP) Gene Expression During Egg Diapause and Its Multi-Gene Organization in the Silkworm Bombyx mori

  • Sirigineedi, Sasibhushan;Murthy, Geetha N.;Rao, Guruprasada;Ponnuvel, Kangayam M.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.26 no.1
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    • pp.31-40
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    • 2013
  • Paralytic peptide binding proteins (PP-BP) are 30KP proteins that show similarity to ENF binding proteins. The ENF-BP act as active regulators of ENF peptides. ENF peptides are multifunctional insect cytokines. The comparison of gene expression in diapause induced and non-diapause eggs at different time intervals after oviposition showed an upregulation of PP at 18h as well as PP-BP at 12 and 18h after oviposition along with few other genes. The current study has been taken up to investigate the role of PP as well as PP-BP in diapause induction in polyvoltine silkworms and to study the multigene organization of PP-BP in the Bombyx mori genome. The tissue specific expression analysis revealed that, PP-BP is highly expressed in fat body followed by egg and brain while no expression was observed in midgut. The expression levels of PP and PP-BP in diapause and non-diapause eggs from 0h to 48h after oviposition, validated through realtime PCR revealed that PP is highly expressed at 18 and 24h while PP-BP expression is higher at 12 and 18h time intervals suggesting their possible role in diapause induction. The whole genome survey of the PP-BP paralogous sequences revealed a total of 46 B. mori PP-BP homologs that are classified into 3 categories viz., ENF-BP, Typical 30KPs and serine/threonine rich 30KPs. These paralogous sequences are distributed on chromosomes 7, 20, 22 and 24, all 30KP and S/T rich 30KP proteins are present in the same locus of chromosome 20.

Isolation of Lactobacillus plantarum HB1 from Tongchimi and Its Nitrite-Scavenging Effect (동치미로부터 분리된 유산균 Lactobacillus plantarum HB1의 아질산염 소거 효과)

  • 유형재;이선숙;이동석;김한복
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.192-196
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    • 2003
  • To obtain large pools of lactic acid bacteria, a strain was isolated from Tongchimi. Through its sugar fermentation and analysis of 16S rRNA gene, it was identified to be Lactobacillus plantarum HB1. This strain is Gram-positive and catalase-negative. In the range of 1~88 bp in the HB1 16S rRNA gene, the HB1 strain was homologous with other L. plantarum strains by almost 100%, and in the range of the rest 32 bp, the HB1 strain showed considerable variation, compared to other strains. Nitrate which may exist in radish can be easily converted to nitrite. The nitrite interacts with amine, and becomes nitrosamine which may cause stomach cancer. The culture obtained by HB1 strain could eliminate 400 ${\mu}M$ nitrite within 1.5 hr. It is necessary to isolate specific components which are involved in nitrite elimination in the culture and to study on its mechanism.

MiR-99a Inhibits Cell Proliferation and Tumorigenesis through Targeting mTOR in Human Anaplastic Thyroid Cancer

  • Huang, Hou-Gang;Luo, Xi;Wu, Shuai;Jian, Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.12
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    • pp.4937-4944
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    • 2015
  • MicroRNAs (miRNAs) are emerging as critical regulators in carcinogenesis and tumor progression. Recently, miR-99a has been reported as a tumor suppressor gene in various human cancers, but its functions in the context of anaplastic thyroid cancer (ATC) remain unknown. In this study, we reported that miR-99a was commonly downregulated in ATC tissue specimens and cell lines with important functional consequences. Overexpression of miR-99a not only dramatically reduced ATC cell viability by inducing cell apoptosis and accumulation of cells at G1 phase, but also inhibited tumorigenicity in vivo. We then screened and identified a novel miR-99a target, mammalian target of rapamycin (mTOR), and it was further confirmed by luciferase assay. Up-regulation of miR-99a would markedly reduce the expression of mTOR and its downstream phosphorylated proteins (p-4E-BP1 and p-S6K1). Similar to restoring miR-99a expression, mTOR down-regulation suppressed cell viability and increased cell apoptosis, whereas restoration of mTOR expression significantly reversed the miR-99a antitumor activity and the inhibition of mTOR/p-4E-BP1/p-S6K1 signal pathway profile. In clinical specimens and cell lines, mTOR was commonly overexpressed and its protein levels were statistically inversely correlated with miR-99a expression. Taken together, our results demonstrated for the first time that miR-99a functions as a tumor suppressor and plays an important role in inhibiting the tumorigenesis through targeting the mTOR/p-4E-BP1/p-S6K1 pathway in ATC cells. Given these, miR-99a may serve as a novel prognostic/diagnostic and therapeutic target for treating ATC.

Environments of Post-Glacial Age at Sorori, Reconstructed by the Species of Excavated Woods (청원 소로리 후빙기 시대의 환경 -출토목재의 수종분석을 중심으로-)

  • 박원규;김수철;이융조
    • The Korean Journal of Quaternary Research
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    • v.17 no.1
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    • pp.1-6
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    • 2003
  • One hundred fifty six pieces of woods were excavated at the muddy sand layer (post-glacial age: about 8,800 bp) above the upper peat layer from Sarori, Chungwongun, Korea in the central Korea. Due to the deteriorated structure of peat woods, all samples were embedded in PEG(polyethylene glycol) 2000 and then sectioned using a rotary microtome. Only two species were found; Alnus spp.(95%) and Ulmus spp.(5%). No conifers were found. Species composition indicates that the climate condition of central Korean peninsula around 8,800 bp was little wetter condition. The sampled region at the Sarori appeared to be a swamp or riverside.

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Interaction of Ras-GTPase-activating Protein SH3 Domain-binding Proteins 2, G3BP2, With the C-terminal Tail Region of KIF5A (Ras-GTPase-activating protein SH3 domain-binding proteins 2, G3BP2와 KIF5A C-말단 꼬리 영역과의 결합)

  • Jeong, Young Joo;Jang, Won Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1191-1198
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    • 2017
  • Vesicles and organelles are transported along microtubule and delivered to appropriate compartments in cells. The intracellular transport process is mediated by molecular motor proteins, kinesin, and dynein. Kinesin is a plus-end-directed molecular motor protein that moves the various cargoes along microtubule tracks. Kinesin 1 is first isolated from squid axoplasm is a dimer of two heavy chains (KHCs, also called KIF5s), each of which is associated with the light chain (KLC). KIF5s interact with many different binding proteins through their carboxyl (C)-terminal tail region, but their binding proteins have yet to be specified. To identify the interacting proteins for KIF5A, we performed the yeast two-hybrid screening and found a specific interaction with Ras-GTPase-activating protein (GAP) Src homology3 (SH3)-domain-binding protein 2 (G3BP2), which is involved in stress granule formation and mRNA-protein (mRNP) localization. G3BP2 bound to the C-terminal 73 amino acids of KIF5A but did not interact with the KIF5B, nor the KIF5C in the yeast two-hybrid assay. The arginine-glycine-glycine (RGG)/Gly-rich region domain of G3BP2 is a minimal binding domain for interaction with KIF5A. However, G3BP1 did not interact with KIF5A. When co-expressed in HEK-293T cells, G3BP2 co-localized with KIF5A and was co-immunoprecipitated with KIF5A. These results indicate that G3BP2, which was originally identified as a Ras-GAP SH3 domain-binding protein, is a protein that interacts with KIF5A.