• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.632-637
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• 2000

This research was undertaken to restore the biological activity of cyclodextrin glucanotransferase (CGTase) of Bacillus macerans origin expressed as inclusion bodies in recombinant Escherichia coli. The optimum concentration of urea used as a denaturant was 8 M. The supplementation of 0.5 M urea into a dialysis buffer increased the refolding efficiency by preventing any protein aggregation. The influence of the protein concentration, temperature, and pH were also investigated. The protein concentration was found to be the most important factor in the refolding efficiency. The optimum temperature was 15-$25^{\circ}C$ and the optimum pH was 6.0. The maximum specific activity of the CGTase refolded under the optimum conditions was 92.2 U/mg, corresponding to 72% of the native CGTase. A comparison of the secondary structure between the native and the refolded CGTase showed that the relative ratio of the $\alpha$-helix content in the native to the refolded CGTase was 1:0.82.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.579-585
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• 2007

Formation of inclusion bodies is usually observed when foreign proteins are overexpressed in E. coli. The formation of inclusion bodies might be prevented by lowering the rate of protein synthesis, and appropriate regulation of the protein expression rate may lead to the soluble expression. In this study, human growth hormone (rhGH) was expressed in a soluble form by slowing down the protein synthesis rate, which was controlled in the transcriptional and translational levels. The transcriptional level was controlled by the regulation of the amount of RNA polymerase specific to the promoter in front of the rhGH gene. For lowering the rate of translation, the T7 transcription terminator-deleted vector was used to synthesize the longer mRNA of the target gene because the longer mRNA is expected to reduce the availability of tree ribosomes. In both methods, the percentage of soluble expression increased when the expression rate slowed down, and more than 93% of rhGH expressed was a soluble form in the T7 transcription terminator-deleted expression system.

• BLOOD RESEARCH
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• 제53권4호
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• pp.265-265
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• 2018

• 대한수의학회지
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• 제21권1호
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• pp.41-43
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• 1981

The occurrence of inclusion body hepatitis was confirmed for the first time in Korea from chickens submitted for diagnosis to this Institute from a farm located in the vicinity of Anyang. The chickens showed no specific clinical signs except for moderate anemic conditions. At autopsy, livers were swollen and mottled with numerous stellate subcapsular hemorrhages and necrotic foci. Severe nephritis and catarrhal enteritis were also seen. The most notable microscopic changes were seen in the liver. These included eosinophilic intra-nuclear inclusion bodies in the hepatocyes, massive hemorrhages and necrosis and fatty changes in the liver parenchema.

• Food Science and Biotechnology
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• 제17권2호
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• pp.219-227
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• 2008

Bacillus thuringiensis was isolated as a pathogen of the sotto disease of silkmoth larvae about a hundred years ago. Since then, this bacterium has attracted attentions of not only insect pathologists but also many other scientists who are interested in its strong and specific insecticidal activity. This has led to the recent worldwide development of B. thuringiensis-based microbial insecticides and insect-resistant transgenic plants, as well as a landmark discovery of par asp orin, a cancer cell-specific cytotoxin produced by B. thuringiensis. In this review, we describe examination of interaction between inclusion proteins of B. thuringiensis and brush border membrane of insects using a surface plasmon resonance-based biosensor, identification and characterization of parasporin-4, the latest parasporin produced by the B. thuringiensis A1470 strain, and an effective method for preparing the parasporin-4 from inclusion bodies expressed in the recombinant Escherichia coli cells.

• 한국임상수의학회지
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• 제17권2호
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• pp.502-504
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• 2000

Epitheliocystis in cultured rock firth was examined Epitheliocystis infected gill epithelial cells resulted in the cells enlarging to 20 to 400${\mu}{\textrm}{m}$ in diameter. Key diagnostic feature is a large, granular, basophilic inclusion. filled with coccoid bodies, which occupies virtually the entire cell.

• 한국전자현미경학회:학술대회논문집
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• 한국현미경학회 2003년도 제34차 추계학술대회
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• pp.26-27
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• 2003

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.700-704
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• 1998

Each of the recombinant structural genes of Helicobacter pylori urease, ureA and ureB, was cloned and overexpressed as inclusion bodies. Solubilization and renaturation of the inclusion bodies were carried out, to accelerate the pairing of sulfhydryl groups and the incorporation of nickel ions, which would lead to the native structure with high enzyme activity. Rates of urea hydrolysis were monitored as an indication of in vitro activation of renatured ureases. The activation of the apoprotein using 1 mM nickel ion, 100 mM sodium bicarbonate and a 10:1 ratio of reducing power resulted in a weak urease activity (about 11% of the native urease activity encoded by pTZ 19R/ure-l). When a sparse matrix screen method originally discovered for the crystallization of proteins was used, the activity increased higher than that obtained using glutathione. The effect of polyethylene glycol (PEG) on the activity was noticeable, giving two-fold increase in the specific activity (about 11 U/mg of protein corresponding to 22% of the native urease activity encoded by pTZ19R/ure-1).

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.590-600
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• 2003

The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

• 대한수의학회지
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• 제31권3호
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• pp.295-302
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• 1991

From 1988 to 1989, 8 strains of canine parvovirus-2(CPV-2) were isolated from the fecal specimens from the dogs that were clinically diagnosed as canine parvoviral enteritis in the veterinary hospitals located in the regions of Taejeon and Chungbuk province. The biological and physicochemical properties for the isolates were studied. Among 62 fecal samples collected from the dogs with enteric diseases, 24(38.7%) showed the haemagglutinating activity to porcine erythrocyte ranging from 16 to 16,384 of HA titers. In cytopathological studies with CRFK cells, intranuclear inclusion bodies were observed in all of eight specimens with the high HA titer over 1,000, of which three specimens showed cytoplasmic inclusions concurrently with the intranuclear inclusion bodies. It was found that the isolates revealed the highest haemagglutinating activity with porcine erythrocytes and the relatively lower haemagglutination titers with the erythrocytes from cat and rabbit. None of erythrocytes from the other animals reacted with the isolates. By the cross-haemagglutination inhibition test for the isolates with the reference viruses and sera, the isolates were evidently identified as the strains of CPV-2. In physicochemical property test, the isolates were stable in lipid solvent, pH and heat treatment at $56^{\circ}C$ for 30 min, and showed the virus particle size less than 25 nm, containing a DNA genome.