• Title/Summary/Keyword: C1-2 fusion

Search Result 544, Processing Time 0.029 seconds

Dynamics of OH Production in the Reaction of O(1D2) with Cyclopropane

  • Jang, Sungwoo;Jin, Sung Il;Kim, Hong Lae;Kim, Hyung Min;Park, Chan Ryang
    • Bulletin of the Korean Chemical Society
    • /
    • v.35 no.6
    • /
    • pp.1706-1712
    • /
    • 2014
  • The OH($X^2{\Pi}$, ${\upsilon}^{\prime\prime}=0,1$) internal state distribution following the reaction of electronically excited oxygen atom ($O(^1D_2)$) with cyclo-$C_3H_6$ has been measured using laser-induced fluorescence, and compared with that following the reaction of $O(^1D_2)$ with $C_3H_8$. The overall characteristics of the OH internal energy distributions for both reactions were qualitatively similar. The population propensity of the ${\Pi}(A^{\prime})$ ${\Lambda}$-doublet sub-level suggested that both reactions proceeded via an insertion/elimination mechanism. Bimodal rotational population distributions supported the existence of two parallel mechanisms for OH production, i.e., statistical insertion and nonstatistical insertion. However, detailed analysis revealed that, despite the higher exoergicity of the reaction, the rotational distribution of the OH following the reaction of $O(^1D_2)$ with $C_3H_8$ was significantly cooler than that with cyclo-$C_3H_6$, especially in the vibrational ground state. This observation was interpreted as the effect of the flexibility of the insertion complex and faster intramolecular vibrational relaxation (IVR).

Identification of a Cellular Protein Interacting with Murine Retrovirus Gag Polyproteins

  • Choi, Wonja
    • Journal of Microbiology
    • /
    • v.34 no.4
    • /
    • pp.311-315
    • /
    • 1996
  • The retroviral Gag polyprotein directs the assembly of virion particles and plays an important role in some events after entry into a host cell. The Gag polyprotein of a virus mixture is responsible for inducing murine acquired immunodeficiency syndrome (MAIDS) when injected into susceptible strains of mice. In order to identify the host cellular proteins which interact with the MAIDS virus Gag proteins and possibly mediate the function of the Gag proteins, mouse T-cell leukemic cDNA expression library was screened using the yeast GAL4 two hybrid system. Of 11 individual positive clones, the clone Y1 was selected for the study of protein-protein interaction. Its DNA sequence revealed that it was an exact match to the murine SH3 domain-containing protein SH3P8. It is expressed as 2.4 kbp transcripts in testis at higher levels and in various tissues tested at lower levels. Glutathione S-transferase-Y1 fusion protein binds tightly to $Pr60^{def-gag}$ as well as $Pr65^{eco-gag}$.

  • PDF

The Fission Yeast Gene Encoding Monothiol Glutaredoxin 5 Is Regulated by Nitrosative and Osmotic Stresses

  • Kim, Hong-Gyum;Park, Eun-Hee;Lim, Chang-Jin
    • Molecules and Cells
    • /
    • v.20 no.1
    • /
    • pp.43-50
    • /
    • 2005
  • Glutaredoxin (Grx) is a small, heat-stable redox protein acting as a multi-functional glutathione (GSH)-dependent disulfide oxidoreductase. We have cloned the monothiol Grx5 gene from the genomic DNA of the fission yeast Schizosaccharomyces pombe. It has 1,904 bp, with one intron, and encodes a putative protein of 146 amino acids with a molecular mass of 16.5 kDa. Recombinant Grx5 produced functional Grx in S. pombe cells. NO-generating sodium nitroprusside (SNP, 1.0 and 2.0 mM) and potassium chloride (KCl, 0.2 and 0.5 M) increased the synthesis of ${\beta}$-galactosidase from a Grx5-lacZ fusion gene, and transcription of Grx5 was also enhanced by SNP and KCl. Synthesis of ${\beta}$-galactosidase from the Grx5-lacZ fusion was lower in Pap1-negative TP108-3C cells than in wild type KP1 cells, and when Pap1 was overproduced in KP1 cells, the level of ${\beta}$-galactosidase increased. We also found that Pap1 is involved in the induction of Grx5 by SNP and KCl. S. pombe Grx5 may play a crucial role in responses to nitrosative and osmotic stresses.

C-terminal Fusion of EGFP to Pneumolysin from Streptococcus pneumoniae modified its Hemolytic Activity (Streptococcus pneumoniae가 생산하는 pneumolysin의 EGFP 융합으로 인한 용혈활성 변화)

  • Chung, Kyung Tae;Lee, Jae Heon;Jo, Hye Ju
    • Journal of Life Science
    • /
    • v.28 no.1
    • /
    • pp.99-104
    • /
    • 2018
  • Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.

Culture Characteristics on the Activity of ${\gamma}-Glutamyltranspeptidase{\;}({\gamma}-GTP)$ by Bacillus subtilis Fusant (배양조건이 Bacillus subtilis 융합주의 ${\gamma}-Glutamyltranspeptidase{\;}({\gamma}-GTP)$ 활성에 미치는 영향)

  • 김관필;김성호;정낙현
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.30 no.3
    • /
    • pp.395-402
    • /
    • 2001
  • A fusant FG-21 was selected on the basis of higher ${\gamma}-GTP$ activity following fusion process between SM-2 and SM-10 of Bacillus subtilis mutants. ${\gamma}-GTP$ activity of the mutant FG-21 was increased up to 612 U/mL when grown for 36 hr at $37^{\circ}C$ in culture media containing 1% glycerol 1% glycerol, 1% peptone, 0.1% citric acid, 5 mM $K_2HPO_4$, 1 mM $FeCl_3$, 1 mM $MgCl_2$, 1 mM $NH_4Cl$, pH 7.0. In fusnat FG-21, the ratio of protein to total sugar contents for biopolymer A was 38 to 59. for biopolymer B from parental strains it was 19 to 78. Fructose contents determined by HPLC were $573.7\;\mu\textrm{g}/mg\;and\;764.4\;\mu\textrm{g}/mg$ for biopolymer A and B, respectively. And glutamic acid content were $163.7\;\mu\textrm{g}/mg\;and\;94.6\;\mu\textrm{g}/mg$ for biopolymer A and B, respectively. In fusant FG-21, the ratio of fructose to glutamic acid contents for biopolymer A was 78 to 22. For biopolymer B from parental strains it was 89 to 11.

  • PDF

Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
    • /
    • v.28 no.1
    • /
    • pp.21-27
    • /
    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).

8-port Coupled Transmission Line Modeling of KSATR ICRF Antenna and Comparison with Measurement (커플링이 고려된 KSTAR ICRF 안테나의 8포트 전송선 회로 모델링 및 측정 결과 비교)

  • Kim, S.H.;Wang, S.J.;Hwang, C.K.;Kwak, J.G.
    • Journal of the Korean Vacuum Society
    • /
    • v.19 no.1
    • /
    • pp.72-80
    • /
    • 2010
  • It is very important to predict and analyze the change of voltage and current distribution of current strap, abnormal voltage distribution of transmission line and resonance phenomenon by coupling between current straps for more stable operation of ICRF system. In this study, to understand those phenomena by coupling, 8-port coupled transmission line model is completed by appling S-parameter measured in the prototype KSTAR ICRF antenna to the model. The determined self-inductance, mutual-inductance and capacitance of antenna straps are shown to be lower than that calculated from 2D approximate model due to finite length of strap. The coupled transmission line model of current strap will be utilized to the operation of ICRF system of KSTAR in the future.

A Study on the production of Monoclonal Antibodies against Rhodotrodula rubra (효모양진균의 단크론 생성)

  • Ryeom K.;Kim S. C.;Lee J. H.
    • Journal of environmental and Sanitary engineering
    • /
    • v.4 no.1 s.6
    • /
    • pp.17-28
    • /
    • 1989
  • Having studied the production of monoclonal antibodies for developing a diagnosis medicine which shall be detected by a high-sensitivity test by using Rhodotorula rubra as a fungi-host which had been extracted through biochemical tests and follow-up examinations on Yeast-like fungi obtained from pulmonary tissues of pulmonary tuberculosis patients who had been in Kong ju National Tuberculosis Hospital from Jun. to Dec. in 1987, I. have gained such results as follows: 1. The fusion rate was influenced by feeder cell layers, cell density and time required to the cell fusion with cells in myelona subculture. 2. The fusion rate did not show any significant difference when the cell was applyed with two molecular weights, i.e., 1500 and 4000, of polyethylene glycol. 3. Fused cells after the addition of HAT selection media were bright and round, whereas unfused myelona cells and spleen cells were shrunk and granulated. 4. The cell fusion rate turned out to be about $57.2\%$(150 wells / 264 wells). 5. $10\%$(15 wells / 150 wells) of the positive reaction was detected in monoclonal antibody screening. 6. The titer which had reacted positively to Rhodotorula rubra fungal-host was 800 times in density after the gradual dilution of the produced monoclonal antibodies with Indirect ELISA method. 7. The Strongest specific reaction came out after the peroxidase labelled anti-human Immunogobulin had been applyed to Rhodotorula rubra for activating its nature after making drift with Carbonate-bicarbonate buffer (pH 9.6) and drying completely.

  • PDF

Production of Nucleocapsid Protein of Newcastle Disease Virus in Escherichia coli and its Assembly into Ring-and Nucleocapsid-like Particles

  • Kho, Chiew-Ling;Tan, Wen-Siang;Khatijah Yusoff
    • Journal of Microbiology
    • /
    • v.39 no.4
    • /
    • pp.293-299
    • /
    • 2001
  • The nucleocapsid(NP) protein of Newcastle disease virus (NDV) and its derivative (NP$\sub$cfus)containing the myc region and six histidine residues fused to its C-terminus were pcpressed aboundantly in Escherichia coli. The proteins were purified by sucrose gradient centrifugation. Both the NP and NP$\sub$cfus/ proteins self-assem- bled into ring-like particles stacked together to from nucleocapsid-like structure which are heterogeneous in length with a diameter of 20${\pm}$2 nm and central holow of 5${\pm}$1 nm. Only a very small amount of the monomers in the particles was linked by inter-molecular disulfide bonds. Fusion of the C-terminal end to 29 amino acids inclusive of the myc epitope and His tag did not impair ring assembly buy inhibited the formation of the long herringbone structures. Immunogold lableing of the particles with the anti-myc antibody showed that the C-terminus of the NP$\sub$cfus/ protein is exposed on the surface of these ring-like particles.

  • PDF

Effect of specimen size on fracture toughness of reduced activation ferritic steel (JLF-l) (저방사화 철강재 (JLF-1)의 파괴인성에 미치는 시험편 크기의 영향)

  • Kim, Dong-Hyun;Yoon, Han-Ki;Park, Won-Jo;Katoh, Y.;Kohyama, A.
    • Proceedings of the Korea Committee for Ocean Resources and Engineering Conference
    • /
    • 2003.05a
    • /
    • pp.300-305
    • /
    • 2003
  • Reduced activation ferritic (JLF-1) steel is leading candidates for blanket/first-wall structures of the D-T fusion reactor. In fusion application, structural materials will suffer effects of repeated changes of temperature. Therefore, the data base of tensile strength and fracture toughness at operated temperature $400^{\circ}C$ are very important. Fracture toughness ($J_{IC}$) and tensile tests were carried out at room temperature and elevated temperature ($400^{\circ}C$). Fracture toughness tests were performed with two type size to investigate the relationship between the constraint effect of a size and the fracture toughness resistance curve. As the results, the tensile strength and the fracture toughness values of the JLF-1 steel are slightly decreased with increasing temperature. The fracture resistance curve increased with increasing plane size and decreased with increasing thickness. The fracture toughness values of JLF-1 steel at room temperature and at $400^{\circ}C$ shows an excellent fracture toughness ($J_{IC}$) of about $530kJ/m^2\;and\;340kJ/m^2$, respectively.

  • PDF