• Title/Summary/Keyword: P13

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Function and Oligomerization Study of the Leucine Zipper-like Domain in P13 from Leucania separata Multiple Nuclear Polyhedrosis Virus

  • Du, Enqi;Yao, Lunguang;Xu, Hua;Lu, Songya;Qi, Yipeng
    • BMB Reports
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    • v.40 no.2
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    • pp.232-238
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    • 2007
  • The p13 gene is uniquely present in Group II nucleopolyhedroviruses (NPVs) and some granuloviruses, but not in Group I NPVs. p13 gene was first described by our laboratory in Leucania separatamultiple nuclear polyhedrosis virus (Ls-p13) in 1995. However, the functions of Ls-P13 and of its homologues are unknown. When Ls-p13 was inserted into Autographa californica nucleopolyhedrovirus, a Group I NPV, polyhedra yield was inhibited. However, this inhibition was prevented when the leucine zipper-like domain of Ls-p13 was mutated. To determine the cause of this marked difference between Ls-P13 and leucine zipper mutated Ls-P13 (Ls-P13mL), oligomerization and secondary structure analyses were performed. High performance liquid chromatography and yeast two-hybrid assays indicated that neither Ls-P13 nor Ls-P13mL could form oligomers. Informatics and circular dichroism spectropolarimetry results further indicated marked secondary structural differences between Ls-P13 and Ls-P13mL. The LZLD of Ls-P13 has two extended heptad repeat units which form a hydrophobic surface, but it is short of a third hydrophobic heptad repeat unit for oligomerization. However, the mutated LZLD of Ls-P13mL lacks the above hydrophobic surface, and its secondary structure is markedly different. This difference in its secondary structure may explain why Ls-P13mL is unable to inhibit polyhedra yield.

p13 from group II baculoviruses is a killing-associated gene

  • Lu, Nan;Du, Enqi;Liu, Yangkun;Qiao, Hong;Yao, Lunguang;Pan, Zishu;Lu, Songya;Qi, Yipeng
    • BMB Reports
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    • v.45 no.12
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    • pp.730-735
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    • 2012
  • p13 gene was first described in Leucania separata multinuclear polyhedrosis virus (Ls-p13) several years ago, but the function of P13 protein has not been experimentally investigated to date. In this article, we indicated that the expression of p13 from Heliothis armigera single nucleocapsid nucleopolyhedrovirus (Ha-p13) was regulated by both early and late promoter. Luciferase assay demonstrated that the activity of Ha-p13 promoter with hr4 enhancer was more than 100 times in heterologous Sf9 cells than that in nature host Hz-AM1 cells. Both Ls-P13 and Ha-P13 are transmembrane proteins. Confocal microscopic analysis showed that both mainly located in the cytoplasm membrane at 48 h. Results of RNA interference indicated that Ha-p13 was a killing-associated gene for host insects H. armigera. The AcMNPV acquired the mentioned killing activity and markedly accelerate the killing rate when expressing Ls-p13. In conclusion, p13 is a killing associated gene in both homologous and heterologous nucleopolyhedrovirus.

Characterization of the Membrane-bound Adenosine Triphosphatase from Corn Roots (옥수수 뿌리로부터 분리한 Membrane-bound ATPase의 특성에 관한 연구)

  • Moon, Hye Yeon;Kwang Soo Roh;Woong Seop Sim
    • Journal of Plant Biology
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    • v.24 no.4
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    • pp.171-179
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    • 1981
  • The membrane-bound ATPases were separated on sucrose gradient from corn roots and characterized by pH optima, sensitivity to monovalent salt, Km and Vmax. The pH optima for the activity of all the ATPases associated with 13, 000g pellet and 13, 000~80, 000g pellet were 5 and 9, respectively. The ATPases in Fractions B and C of the 13, 000 g pellet were more active at pH 5 than pH 9. While, in the case of Fractions D, E and F, they were reverse. The activities of the ATPase in Fractions A and C of the 13, 000~80, 000 g pellet were greater at pH 5 than pH 9. On the other hand, the ATPases in Fractions B, D, E, and F were more active at pH 9 than pH 5. The optimum concentraction of ATP for the assay was about 3 to 5 mM. The Km's for the membrane-bound ATPases in 13, 000g pellet and in 13, 000~80, 000 g pellet were 0.25 mM. While Vmax values for 13, 000g pellet were from 8.0 to 12.5 $\mu$M Pi/mg protein/hr. according to pH values, those for 13, 000~80, 000 g pellet were from 35.7 to 55.6 $\mu$M Pi/mg protein/hr. Activities of the membrane-bound ATPases in both 13, 000 g pellet and 13, 000~80, 000 g pellet were stimulated with increasing the concentration of $K^+$.

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FUNGAL EXTRACELLULAR POLYSACCHARIDES INVOLVED IN RECYCLING OF METABOLITES AND OSMOTOLERANCE OF PENICILLIUM FELLUTANUM : APPLICATION OF $^{13}$ C-NMR SPECTROSCOPY FOR THE STUDY ON FUNGAL PHYSIOLOGY AND METABOLISM

  • Park, Yong-Il;Gander, John.-E.
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.208-213
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    • 2000
  • Penicillium fellutanum produces a phosphorylated, choline-containing extracellular peptido-polysaccharide, peptidophosphogalactomannan (pPxGM) (8). The $\^$13/C-methyl labeled pPxGM ([methyl-$\^$13/C]pPxGM) was prepared from the cultures supplemented with L-[methyl-$\^$13/C]methionine or [2-$\^$13/C]glycine and was used as a probe to monitor the fate of phosphocholine in this polymer. Addition of purified [methyl-$\^$l3/C]pPxGM to growing cultures in low phosphate medium resulted in the disappearance of [methyl-$\^$13/C]phosphocholine and -N,N'-dimethyl-phosphoethanolamine from the added [methyl-$\^$13/C]pPxGM. Two $\^$l3/C-methyl-enriched cytoplasmic solutes, choline-O-sulfate and glycine betaine, were found in mycelial extracts, suggesting that phosphocholine-containing extracellular pPxGM of P.fellutanum is a precursor of intracellular choline-O-sulfate and glycine betaine and thus of phosphatydilcholine (l0). $\^$13/C-Methyl-labeled cells grown in 3 M NaCl-containing medium showed 2.6- and 22-fold more accumulation of $\^$13/C-methyl labeled choline-O-sulfate and glycine betaine, respectively, originated from the extracellular [$\^$13/C-methyl]pPxGM than those grown without added NaCl. The results suggest that, in addition to glycerol and erythritol, glycine betaine and choline-O-sulfate and thus choline are also osmoprotectants and hence that pPxGM is involved in osmotolerance of this fungus (11). Taken collectively, the $\^$l3/C- and $\^$31/P-NMR analyses of cytosolic solute pools and structural modulation of extracellular pPxGM corresponding to environmental stimuli in P. fellutanum, provided evidence that pPxGM is involved in cellular choline metabolism, osmotolerance, and recycling of metabolites.

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A Study on Basic Investigation for Quality Improvement of Raw-Silk A Study on the Dissolving Behavior of Sericin in the Cocoon Shell and Pupa Protein (Bombyx mori L.) (Part III) (생사 품위향상을 위한 기초조사 연구 견층 Sericin과 용체 단백질의 용해거동에 관한 연구(III))

  • 임영우
    • Journal of Sericultural and Entomological Science
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    • v.17 no.2
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    • pp.161-170
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    • 1975
  • This study is to investigate the content of crude protein intruded in the sericin of cocoon shell and pupa by treatment of buffer solution (pH 1 to pH 13) 20 ml per 1 gram for 30 and 60 minutes at 30$^{\circ}C$, 60$^{\circ}C$ and 100${\pm}$2$^{\circ}C$, respectively. The results obtained are summarized as follows. 1. The quantity of total crude protein obtained from cocoon shell and pupa by treatment during 30 minutes at 30$^{\circ}C$ was dissolved as the largest quantity of 11.874 mg/g at pH 1 and l5.93mg/g at pH 13, but dissolved the smallest quantity 1.75g/g at pH 5 as known. and tile quantity of crude protein treated for 60 minutes is 13.437mg/g at pH 1 and 22.50mg/g at pH 13. Also, the smallest quantity is 2. 813mg/g at pH 5 as known. 2. By the treatment during 30 minutes at 60$^{\circ}C$, the dissolved largest quantity was 13.12mg/g at pH 1 and 21.875 mg/g at pH 13, but the smallest quantity is 2.375mg/g at pH 5 as known After treatment for 60 minutes at 60$^{\circ}C$, the dissolved largest quantity was 17.500 mg/g at pH 1 and 31.56mg/g at pH 13, bu the smallest quantity is 3.849 mg/g at pH 5. 3. The dissolved crude protein from the cocoon shell and pupa by treatment for 30 minutes at 100${\pm}$2$^{\circ}C$ was the largest quantity of 147.000mg/g at pH 1 and 398. 125mg/g at pH 13, but the smallest quantity is 75.00mg/g at pH 5 as known. After treatment for 60 minutes at 100${\pm}$2$^{\circ}C$, the largest quantity was 253.76 mg/g at pH 1 and 460.625mg/g at pH 13, but the smallest quantity is 139.375mg/g at pH 5 as known. 4. The dissolved crude protein from the cocoon shell and pupa was not different in quantity by treatment at 30$^{\circ}C$ or 60$^{\circ}C$. But dissolved crude protein was large quantity from cocoon shell more than pupa, as known. 5. The treatment of cocoon shell and pupa during 60 minutes at 100${\pm}$20$^{\circ}C$ was increased to the dissolved largest quantity of crude protein of 19.20% at pH 1 and 22. 18% at pH 13 from the cocoon shell and 6. 12% at pH, an d 23.87% at pH 13 from the pupa. But dissolved crude protein was increased to the larger quantity from pupa more than cocoon shell.

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Molecular Cloning and Expression of a Xylanase Gene from Thermophilic Alkalophilic Bacillus sp. K-17 in Escherichia coli (고온, 호알칼리성 Bacillus sp. K-17 Xylanase 유전자의 Escherichia coli 에의 클로닝 및 발현)

  • Sung, Nack-Kie;Chun, Hyo-Kon;Shim, Ki-Hwan;Kang, In-Soo;Teruhiko Akiba
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.178-182
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    • 1989
  • A gene coding for a xylanase of thermophilic alkalophilic Bacillus sp. K-17 was cloned in Escherichia coli C600 with pBR322. Plasmid pAXl13 was isolated from a transformant producing xylanase, and the xylanase gene was located in a 4.3 Kb HindIII fragment. Biotinylated pAXl13 hybridized to a 4.3 Kb HindIII fragment from chromosomal DNA of thermophilic alkalophilic Bacillus sp. K-17. The xylanase activity was observed in the extracellular curture fluid of E. coli carrying pAXl13. The pAXl13-encoded xylanase had the same enzymatic properties as those of xylanase I produced by thermophilic alkalophilic Bacillus sp. K-17.

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Construction of a Lactococcal Shuttle/Expression Vector Containing a $\beta$-Galactosidase Gene as a Screening Marker (선별마커로써 $\beta$-Galactosidase 유전자를 포함한 Lactococcus용 셔틀/발현 벡터 제조)

  • Han Tae Un;Jeong Do-Won;Cho San Ho;Lee Jong-Hoon;Chung Dae Kyun;Lee Hyong Joo
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.241-247
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    • 2005
  • A new lactococcal shuttle/expression vector for lactococci, pWgal13T, was constructed using a $\beta$-galactosi-dase gene (lacZ) from Lacfococcus lactis ssp. lactis ATCC 7962 as a screening marker. The pWgal 13T was introduced into Escherichia coli DH5a and L. lactis MG1363, and was easily detected by the formation of blue colonies on a medium containing X-gal without any false transformants. Also, the quantitatively lacZ activity of pWgal13T was measured in L. lactis ssp. cremoris MG1363, and was found to be four times higher than that of L. lactis ssp. lactis ATCC7962 grown on a medium containing glucose, which shows that the lacZ gene of pWgal13T can be used for the efficient screening of L. lactis on general media. The pWgal13T was equipped with a lactococcal replicon of pWV01 from L. lactis Wg2, the new promoter P13C from L. lactis ssp. cremoris LM0230, multiple cloning sites, and a terminator for the expression of a relevant gene. The vee-tor pWgal13T was used for the expression of the EGFP gene in E. coli and L. lactis. These results show that the lactococcal expression/shuttle vector constructed in the present study can be used for the production of foreign proteins in E. coli and L. lactis.

A Study of Sex Difference by Chemical Determination of Dentin in Human Teeth (치아경조직의 화학적 판정에 의한 성차 연구)

  • 장세활;김종열
    • Journal of Oral Medicine and Pain
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    • v.8 no.1
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    • pp.127-142
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    • 1983
  • This thesis was to investigate sex defferencd by employing the methodes of Bernadskij's and it's modification by means of the evaluation of pH of dentin in 162 extracted teeth. The obtained were as follows: 1. In 72% of all experinments, pH of human dentin substance was higher in female than in male. Mean pH was 13.175±0.031 in male and 13.245±0.030 in female. 2. After comparative studies on each pH in anatomical region of tooth arrangement, the sex difference of pH was most obviously showed in molars. 3. In pH change according to ages, the 30's was the highest in male and the 20's in female, also pH showed decresing tendency by the increasing of age in both male and female. 4. There was no pH difference in anatomical regions of tooth arrangment. 5. Mean pH was 13.282±0.013 in under 50 years of age and 13.282±0.022 over 50. It proved that pH was higher in under 50 years of age than in over 50.

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Evaluation of Perfusion and Image Quality Changes by Reconstruction Methods in 13N-Ammonia Myocardial Perfusion PET/CT (13N-암모니아 심근관류 PET/CT 검사 시 영상 재구성 방법에 따른 관류량 변화와 영상 평가)

  • Do, Yong Ho;Lee, Hong Jae;Kim, Jin Eui
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.69-75
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    • 2014
  • Purpose: The aim of this study was to evaluate changes of quantitative and semi-quantitative myocardial perfusion indices and image quality by image reconstruction methods in $^{13}N$-ammonia ($^{13}N-NH_3$) myocardial perfusion PET/CT. Materials and Methods: Data of 14 (8 men, 6 women) patients underwent rest and adenosine stress $^{13}N-NH_3$ PET/CT (Biograph TruePoint 40 with TrueV, Siemens) were collected. Listmode scans were acquired for 10 minutes by injecting 370MBq of $^{13}N-NH_3$. Dynamic and static reconstruction was performed by use of FBP, iterative2D (2D), iterative3D (3D) and iterative TrueX (TrueX) algorithm. Coronary flow reserve (CFR) of dynamic reconstruction data, extent(%) and total perfusion deficit (TPD) (%) measured in sum of 4-10 minutes scan were evaluated by comparing with 2D method which was recommended by vendor. The image quality of each reconstructed data was compared and evaluated by five nuclear medicine physicians through a blind test. Results: CFR were lower in TrueX 18.68% (P=0.0002), FBP 4.35% (P=0.1243) and higher in 3D 7.91% (P<0.0001). As semi-quantitative values, extent and TPD of stress were higher in 3D 3.07%p (P=0.001), 2.36%p (P=0.0002), FBP 1.93%p (P=0.4275), 1.57%p (P=0.4595), TrueX 5.43%p (P=0.0003), 3.93%p (P<0.0001). Extent and TPD of rest were lower in FBP 0.86%p (P=0.1953), 0.57%p (P=0.2053) and higher in 3D 3.21%p (P=0.0006), 2.57%p (P=0.0001) and TrueX 5.36%p (P<0.0001), 4.36%p (P<0.0001). Based on the results of the blind test for image resolution and noise from the snapshot, 3D obtained the highest score, followed by 2D, TrueX and FBP. Conclusion: We found that quantitative and semi-quantitative myocardial perfusion values could be under- or over-estimated according to the reconstruction algorithm in $^{13}N-NH_3$ PET/CT. Therefore, proper dynamic and static reconstruction method should be established to provide accurate myocardial perfusion value.

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Changes of carbon-13 Isotope of Dissolved Inorganic Carbon Within Low-pH CO2-rich Water during CO2 Degassing (pH가 낮은 탄산수의 CO2 탈기에 따른 용존탄소동위원소 변화)

  • Chae, Gitak;Yu, Soonyoung;Kim, Chan Yeong;Park, Jinyoung;Bang, Haeun;Lee, Inhye;Koh, Dong-Chan;Shinn, Young Jae;Oh, Jinman
    • Journal of Soil and Groundwater Environment
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    • v.24 no.3
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    • pp.24-35
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    • 2019
  • It is known that ${\delta}^{13}C_{DIC}$ (carbon-13 isotope of dissolved inorganic carbonate (DIC) ions) of water increases when dissolved $CO_2$ degases. However, ${\delta}^{13}C_{DIC}$ could decrease when the pH of water is lower than 5.5 at the early stage of degassing. Laboratory experiments were performed to observe the changes of ${\delta}^{13}C_{DIC}$ as $CO_2$ degassed from three different artificial $CO_2$-rich waters (ACWs) in which the initial pH was 4.9, 5.4, and 6.4, respectively. The pH, alkalinity and ${\delta}^{13}C_{DIC}$ were measured until 240 hours after degassing began and those data were compared with kinetic isotope fractionation calculations. Furthermore, same experiment was conducted with the natural $CO_2$-rich water (pH 4.9) from Daepyeong, Sejong City. As a result of experiments, we could observe the decrease of DIC and increase of pH as the degassing progressed. ACW with an initial pH of 6.4, ${\delta}^{13}C_{DIC}$ kept increasing but, in cases where the initial pH was lower than 5.5, ${\delta}^{13}C_{DIC}$ decreased until 6 hours. After 6 hours ${\delta}^{13}C_{DIC}$ increased within all cases because the $CO_2$ degassing caused pH increase and subsequently the ratio of $HCO_3{^-}$ in solution. In the early stage of $CO_2$ degassing, the laboratory measurements were well matched with the calculations, but after about 48 hours, the experiment results were deviated from the calculations, probably due to the equilibrium interaction with the atmosphere and precipitation of carbonates. The result of this study may be not applicable to all natural environments because the pressure and $CO_2$ concentration in headspace of reaction vessels was not maintained constant as well as the temperature. Nevertheless, this study provides fundamental knowledge on the ${\delta}^{13}C_{DIC}$ evolution during $CO_2$ degassing, and therefore it can be utilized in the studies about carbonated water with low pH and the monitoring of geologic carbon sequestration.