• Journal of Microbiology
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• v.43 no.3
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• pp.313-318
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• 2005

In this study, we demonstrate the use of a PCR-based method for the detection of the specific genes involved in natural-product biosynthesis. This method was applied, using specifically designed PCR primers, to the amplification of a gene segment encoding for sedo-heptulose 7-phosphate cyclase, which appears to be involved in the biosynthetic pathways of $C_7N$ aminoacyclitol or its keto analogue-containing metabolites, in a variety of actinomycetes species. The sequences of DNA fragments (about 540 bp) obtained from three out of 39 actinomycete strains exhibited a high degree of homology with the sedo-heptulose 7-phosphate cyclase gene, which has been implicated in acarbose biosynthesis. The selective cultivation conditions of this experiment induced the expression of these loci, indicating that the range of $C_7N$ aminoacyclitol or its keto analogue-group natural products might be far greater than was previously imagined. Considering that a total of approximately 20 $C_7N$ aminoacyclitol metabolites, or its keto analogue-containing metabolites, have been described to date, it appears likely that some of the unknown loci described herein might constitute new classes of $C_7N$ aminoacyclitol, or of its keto analogue-containing metabolites. As these metabolites, some of which contain valienamine, are among the most potent antidiabetic agents thus far discovered, the molecular detection of specific metabolite-producing actinomycetes may prove a crucial step in current attempts to expand the scope and diversity of natural-product discovery.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.218-218
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• 2002

Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.

• BMB Reports
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• v.42 no.7
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• pp.418-420
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• 2009

We describe a method for producing polyclonal antibodies against peptide antigen cytochrome P450 1A2 and 3A4 using a tandem repeat of the epitope region and incorporation of proline residue between the repeated sequences. An ELISA assay revealed more efficient generation of polyclonal antibodies to tandem repeat peptide antigens than mono-epitope peptides. The incorporation of proline residues further stimulated antibody production.

• Journal of Plant Biotechnology
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• v.4 no.4
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• pp.171-178
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• 2002

Chlorophyll (Chl) fluorescence imaging was used to investigate the effectiveness of in vivo incorporation methods for two chemicals, 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) and methyl viologen (MV) in rice, a monocot, and cucumber, a dicot, leaves. four different methods (vacuum infiltration, floating, transpiration-aided incorporation through petiole and spraying) were compared, and $F_i$ and $F_v$/$F_m$ were chosen for the imaging of the DCMU- and MV-treated leaves, respectively. The effects of the chemicals in plants were generally heterogeneous over the whole leaf area. Moreover, the effectiveness of the treatment of a chemical in plant leaves was dependent on chemical species, plant species, concentration of the chemical, the treatment method, the duration of the treatment, the existence of light and detergent, etc. In conclusion, we suggest that to achieve the proposed effects of chemicals in plants for an actual experiment, these factors must be considered before the chemical treatment, and the best method for the treatment of the chemicals tested was floating and vacuum infiltration in the dicot and the monocot leaves, respectively, as drawn from Chl fluorescence imaging analysis.

• Bulletin of the Korean Chemical Society
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• v.20 no.6
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• pp.715-719
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• 1999

The incorporation of cobalt into mesoporous molecular sieves MCM-41 and MCM-48 was carried out. Co-PO/MCM41 and Co-PO/MCM48 were prepared using Co(II) acetate solution adjusted to pH = 3.0 with phosphoric acid by the incipient wetness method. Photoacoustic spectroscopy (PAS) was used to study the local environments of Co(II) incorporated into mesopores. The band around 500 nm in PAS of as-prepared Co-PO/MCM41 and Co-PO/MCM48 with Co(II) acetate solution was changed to triplet bands around 600 nm. This could be assigned to the 4 A2(F)-> 4T1(P) transition of Co(II) surrounded tetrahedrally by oxygen ions after calcination. It may be attributable to that the octahedral cobalt species containing phosphate ligands in coordination sphere reacting with framework's silanol groups to be dispersed atomically onto the surface of mesoporous molecular sieves as a tetrahedral species. This is unlike that the Co in Co-Cl/MCM41 and direct-synthesized Co-MCM41 transforms to Co oxide phase upon calcination. Co-PO/MCM41 and Co-PO/MCM48 were stable while treated with water.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.223-223
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• 2006

A Suzuki-coupling route is developed for preparation of ortho-phenylene-bridged Cp/amido complexes, one of which shows higher activity, higher 1-hexene incorporation, and higher molecular weight than the silylene-bridged standard CGC $[Me_{2}Si({\eta}^{5}-Me_{4}Cp)(N^{t}Bu)]TiCl_{2}$.

• Bulletin of the Korean Chemical Society
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• v.25 no.9
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• pp.1366-1370
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• 2004

Controlled pore glass bead was modified with bovine serum albumin (BSA), and glutathione (GSH) was immobilized through three kinds of linkers on top of BSA. Bis(3-sulfo-N-hydroxysuccinimide suberate) sodium salt $(BS^3)$, N-hydroxysuccinimide 3-(2-pyridyldithio)propionate (SPDP), or N-hydroxysuccinimide 4-maleimidobutyrate (GMBS) was introduced into the BSA-bound matrix. Subsequently, GSH was immobilized by addition of thiol side chain into the maleimido moiety, replacing a disulfide group, or formation of an amide group upon releasing 3-sulfo-N-hydroxysuccimide group. It was observed that conjugation methodology played a critical role for activity of the immobilized GSH. SDS-PAGE chromatogram showed that the matrix of glutathione immobilized on BSA through GMBS manifested high selectivity towards glutathione-S-transferase (GST) in cell lysate.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.224-224
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• 2002

Chitin-degrading marine bacterial strain 98CJ11027 was isolated from bryozoa from the coastal area of Cheju Island, Korea, and identified as a member of the genus Vibrio. The molecular mass of the main extracellular chitinase (chitinase I), purified from strain 98CJ11027, was estimated to be 98 kDa. The optimal condition for chitinase I activity is pH 6.0 and 45℃. The activity was inhibited by $Fe^+2$ and$Cu^+2$. Chitinase I displayed the hydrolysis type of chitobiosidase and catalyzed reversed hydrolysis leading to the synthesis of tetraacetylchitotetraose.

• Proceedings of the Polymer Society of Korea Conference
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• 2006.10a
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• pp.159-160
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• 2006

We developed novel catalyst, PHENICS composed of the combination of a cyclopentadienyl group to perform a high catalytic activity and a bulky phenoxy group, which performs the production of high molecular weight polyolefin. The polymerization activity of PHENICS at high temperature is higher than well-known CGC catalyst. PHENICS showed the excellent ability of comonomer incorporation into polymer chain. The obtained copolymer had a high molecular weight. The PHENICS catalyst is also active to the copolymerization of ethylene and several vinyl comonomers such as styrene, norbornen, and conjugated dienes. We will discuss new cocatalysts for PHENICS to improve activity and the ability of molecular weight control.

• Journal of Plant Biology
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• v.36 no.3
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• pp.293-296
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• 1993

Effects of dark treatment and N1-dichlorophenyl-N3-dimethylurea (DCMU) on the desaturation of galactolipids of Dunaliella salina were investigated to see whether light-driven photosynthetic electron transport is involved in in vivo desaturation of galactolipids. The incorporation of radioactive fatty acid precursors ([14C]lauric acid) into galactolipids, mainly composed of prokaryotic molecular species, was most affected among different polar lipid classes by both treatments. The analysis of specific radioactivities of individual galactolipid molecular species revealed that their synthesis was greatly inhibited by the treatments except for eukaryotic molecular species, 18 : 3/ 18 : 3 digalactosyldiacylglycerol, whose desaturation occurs in endoplasmic reticulum.