• Reproductive and Developmental Biology
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• v.34 no.4
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• pp.289-297
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• 2010

Erythropoietin (EPO) is a glycoprotein hormone secreted from primarily cells of the peritubular capillary endothelium of the kidney, and is responsible for the regulation of red blood cell production. We constructed and expressed dimeric cDNAs in Chinease hamster ovary (CHO) cells encoding a fusion protein consisting of 2 complete human EPO domains linked by a 2-amino acid linker (Ile-Asp). We described the activity of dimeric hyperglycosylated EPO (dHGEPO) mutants containing additional oligosaccharide chains and characterized the function of glycosylation. No dimeric proteins with mutation at the $105^{th}$ amino acid were found in the cell medium. Growth and differentiation of the human EPO-dependent leukemiae cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of dHGEPO proteins. MIT assay at 24 h increased due to the survival of F36E cells. The dHGEPO protein migrated as a broad band with an average molecular mass of 75 kDa. The mutant, dHGEPO, was slightly higher than the wild-type (WT) dimeri-EPO band. Enzymatic N-deglycosylation resulted in the formation of a narrow band with a molecular mass twice of that of of monomeric EPO digested with an N-glycosylation enzyme. Hematocrit values were remarkably increased in all treatment groups. Pharmacokinetic analysis was also affected when 2.5 IU of dHGEPO were intravenously injected into the tails of the mice. The biological activity and half-life of dHGEPO mutants were enhanced as compared to the corresponding items associated the WT dimeric EPO. These results suggest that recombinant dHGEPO may be attractive biological and therapeutic targets.

• Journal of Life Science
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• v.14 no.4
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• pp.627-635
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• 2004

In order to distinguish the genetic polymorphism among the olive flounder (Paralichthys olivaceus) obtained from East sea of Jumunjin, aquaculture of Tongyoung and Geoje, and East sea of North Korea, the ND-4 and cytochrome b genes of olive flounder were divided into 5 regions. Each region was analyzed by degenerating gel electrophoresis scanning (DGES), and by subsequent DNA sequencing. The DGES disclosed characteristic DNA polymorphisms in ND-4-2 and ND-4-3 regions of olive flounder, which were also confirmed by the DNA sequencing. The olive flounders obtained from the different marine areas showed DNA mutations in ND-4-2 region (G390A, C402T, and A411G； GenBank： AB028664), and also showed frequent DNA mutations in ND-4-3 region (C515G, C537T, C538T, A567G, G714A, C736T, G756A, A759T, T817C, and T829G), white the cytochrome b gene showed no DNA mutation both in the DGES and DNA sequencing. These data suggest that the ND-4-2 and ND-4-3 regions are candidate loci to distinguish the origin of olive flounder, and that the DGES used in this study provided fast and reliable informations for the genetic polymorphism.

• Journal of KIISE:Software and Applications
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• v.32 no.11
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• pp.1059-1070
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• 2005

For genetic algorithms, the population may get stuck in a local optimum. The population can escape from this after a long duration. This phenomenon is called punctuated equilibrium. The punctuated equilibria observed in nature and computational ecosystems are known to be well described by diffusion equations. In this paper, simple genetic algorithms are theoretically analyzed to show that they can also be described by a diffusion equation. When fitness is the function of unitation, this analysis can be further refined to make the parameters of genetic algorithms appear in this equation. Using theoretical results on the diffusion equation, the duration of equilibrium is shown to be exponential of such parameters as population size, 1/(mutation probability), and potential barrier. This is corroborated by simulation results for bistable potential landscapes with one local optimum and one global optimum.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.12 no.2
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• pp.99-103
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• 2012

Tyrosinemia type I is an autosomal recessive inborn error of tyrosine metabolism that caused a mutation. Clinical symptoms include progressive liver damage with liver failure, coagulopathy, hypophosphataemic rickets, renal tubular dysfunction and a high risk of hepatocellular carcinoma. If left untreated, the affected infants may die from liver failure within the first year of life. PharmacoloIcal therapy with 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC) has offered an effective therapeutic option in addition to dietary restriction of tyrosine and phenylalanine. As prognosis of tyrosinemia type I is improving with early diagnosis and early treatments, it meets the criteria for a condition that would benefit from newborn screening. We report a case of tyrosinemia type I diagnosed by newborn screening and successive biochemical analysis of plasma and urine, treated by dietary restriction and NTBC.

• The Korean Journal of Zoology
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• v.33 no.1
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• pp.6-11
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• 1990

Analysis of genetic structure in Kimpo natural population of Drosophila was carried out by utilizing the deleterious gene on the second chromosome of Drosophila melanogaster. Male flies tested were continuously collected for eight years; in late September 1974 and 1981-1987. The frequency of deleterious gene (lethal plus semilethal) ranged from 27.02% in 1983 to 41.48% in 1987, and the values estimated from the eight years samples are highly signihcent from each other with a homogenety test (X$^2$=52.0157, d.f.=28, P<0.005). Allelic rates ranged from 1.30% in 1981 to 5.03% in 1974. And the effective population size by using the rate of allelism was estimated average at 3, 300 pairs. Elimination rate by homozygous of lethal gene ranged from 0.0004 in 1984 to 0.0019 in 1974, and that is for smaller than mutation rate(0.005) at second chromosome. We suppose that stable frequency (about 20%) lethal genes of D. melanogaster in Kimpo natual population are maintained by invade of P-type mutator factor (P element) versus eliminated in heterozygous and homozygous condition of lethal gene.

• Korean Journal of Veterinary Service
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• v.35 no.3
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• pp.169-174
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• 2012

Japanese encephalitis virus (JEV) is one of the main causes of viral encephalitis in human and animals. For over 30 years, a live attenuated JEV vaccine strain has been used in the veterinary field, and it is required to conduct quality evaluation studies on the commercial vaccines. For the quality control of live attenuated JEV vaccine, we investigated the nucleotide sequence similarity of prME gene derived from five JEV vaccines commercially available in pigs in Korea. The Vero cells infected with JEV vaccines showed specific cytopathic effect, which was characterized by rounding and detached cells. In the phylogenetic analysis, all of the vaccine strains showed a close relationship with the original vaccine seed strain (Anyang 300) and clustered into the genotype 3. In comparison of the nucleotide and deduced amino acid sequences of prME genes with the original strain, all JEV vaccine strains showed high amino acid similarity ranging from 98.9% to 99.5%, but had several point mutations, probably due to high mutation rates of viral RNA polymerase by several virus passages. Even though the current JEV vaccine strains have been maintained and produced for a long period of time, the genetic characterization of them have been rarely changed. However, since the mid 1990's, molecular epidemiology of JEV has been changed sharply from genotype 3 to genotype 1 in Korea, further studies on new vaccine strains to genotype 1 is required for more effective prevention in the field.

• Molecules and Cells
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• v.19 no.1
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• pp.104-113
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• 2005

Large inversions are well characterized in the chloroplast genomes of land plants. In contrast, reports of small inversions are rare and involve limited plant groups. In this study, we report the widespread occurrence of small inversions ranging from 5 to 50 bp in fully and partially sequenced chloroplast genomes of both monocots and dicots. We found that small inversions were much more common than large inversions. The small inversions were scattered over the chloroplast genome including the IR, SSC, and LSC regions. Several small inversions were uncovered in chloroplast genomes even though they shared the same overall gene order. The majority of these small inversions were located within 100 bp downstream of the 3' ends of genes. All had inverted repeat sequences, ranging from 11 to 24 bp, at their ends. Such small inversions form stem-loop hairpin structures that usually have the function of stabilizing the corresponding mRNA molecules. Intra-molecular recombination between the inverted sequences in the stem-forming regions are responsible for generating flip-flop orientations of the loops. The presence of two different orientations of the stem-loop in the trnL-F noncoding region of a single species of Jasminum elegans suggests that a short inversion can be generated within a short period of time. Small inversions of non-coding sequences may influence sequence alignment and character interpretation in phylogeny reconstructions, as shown in nine species of Jasminum. Many small inversions may have been generated by parallel or back mutation events during chloroplast genome evolution. Our data indicate that caution is needed when using chloroplast non-coding sequences for phylogenetic analysis.

• Molecules and Cells
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• v.25 no.3
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• pp.385-389
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• 2008

Septins are a family of filament-forming GTP-binding proteins involved in a variety of cellular process such as cytokinesis, exocytosis, and membrane dynamics. Here we report the biochemical and immunocytochemical characterization of a recently identified mammalian septin, SEPT12. SEPT12 binds GTP in vitro, and a mutation (Gly56 to Asn) in the GTP-binding motif abolished binding. Immunocytochemical analysis revealed that wild-type SEPT12 formed filamentous structures when transiently expressed in Hela cells whereas $SEPT12^{G56A}$ generated large aggregates. In addition, wild-type SEPT12 failed to form filaments when coexpressed with $SEPT12^{G56A}$. We also observed that GTP-binding by SEPT12 is required for interaction with SEPT11 but not with itself.

• Molecules and Cells
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• v.39 no.3
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• pp.211-216
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• 2016

CYP107W1 from Streptomyces avermitilis is a cytochrome P450 enzyme involved in the biosynthesis of macrolide oligomycin A. A previous study reported that CYP107W1 regioselectively hydroxylated C12 of oligomycin C to produce oligomycin A, and the crystal structure of ligand free CYP107W1 was determined. Here, we analyzed the structural properties of the CYP107W1-oligomycin A complex and characterized the functional role of the Trp178 residue in CYP107W1. The crystal structure of the CYP107W1 complex with oligomycin A was determined at a resolution of $2.6{\AA}$. Oligomycin A is bound in the substrate access channel on the upper side of the prosthetic heme mainly by hydrophobic interactions. In particular, the Trp178 residue in the active site intercalates into the large macrolide ring, thereby guiding the substrate into the correct binding orientation for a productive P450 reaction. A Trp178 to Gly mutation resulted in the distortion of binding titration spectra with oligomycin A, whereas binding spectra with azoles were not affected. The Gly178 mutant's catalytic turnover number for the 12-hydroxylation reaction of oligomycin C was highly reduced. These results indicate that Trp178, located in the open pocket of the active site, may be a critical residue for the productive binding conformation of large macrolide substrates.

• Korean Journal of Medicinal Crop Science
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• v.3 no.1
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• pp.50-55
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• 1995

This study was conducted to determine the optimum conditions for induction and different somatic of somatic embryos as well as germination of encapsulated and stored somatic embryos. Somatic embryos was better formed in 1/2X MS medium than full - strength MS medium. 0.1 to 1.0mg/lBA and kinetin promoted shoot differentiation of somatic embryos. Higher concentration tend to inhibit differentiation. IAA affect positively both root and shoot growth. In vitro germination of somatic embryos encapsulated with 2% alginate matrix containing 1/2 MS nutrient medium and $AgNO_3$ 5mg/l was 86%. Storage of somatic embryos was effecive at $5^{\circ}C$ but the germination rate decreased with longer storage period. RAPD analyses with plants regenerated from the somatic embryos showed DNA polymorphism, indicating abolition of primer binding site by point mutation, deletion, or insertion of certain sequences.