• Proceedings of the Korea Society of Poultry Science Conference
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• 1999.11a
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• pp.34-50
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• 1999

A cDNA encoding chicken interferon-gamma (chIFN-${\gamma}$) was amplified from P34, a CD4$^{+}$ T-cell hybridoma by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pUC18. THe sequences of cloned PCR products were determined to confirm the correct cloning. Using this cDNA as probe, chicken genomic library from White Leghorn spleen was screened. Phage clones harboring chicken interferon-gamma (chIFN-${\gamma}$) were isolated and their genomic structure elucidated. The chIFN-${\gamma}$ contains 4 exons and 3 introns spanning over 14 kb, and follows the GT/AG rule for correct splicing at the exon/intron boundaries. The four exons encode 41, 26, 57 and 40 amino acids, respectively, suggesting that the overall structure of IFN-${\gamma}$ is evolutionairly conserved in mammalian and avian species. The 5’-untranslated region and signal sequences are located in exon 1. Several AT-rich sequences located in the fourth exon may indicate a role in mRNA turnover. The 5’-flanking region contains sequences homologous to the potential binding sites for the mammalian transcription factors, activator protein-1(AP-1) activator protein-2(AP-2) cAMP-response element binding protein(CREB), activating transcription factor(ATF), GATA-binding fator(GATA), upstream stimulating factor(USF), This suggests that the mechanisms underlying transcriptional regulation of chicken and mammalian IFN-${\gamma}$ genes may be similar.r.

• Journal of Microbiology
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• v.34 no.4
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• pp.311-315
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• 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 $Pr65^{eco-gag}$.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1568-1579
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• 2001

This study is aimed at evaluating the effect of Aspergillus oryzae fermentation extract (AFE) on corn silage fermentation characteristics. Trial included two groups of treatments, with or without AFE inclusion in corn ensilage. Sixty corn silage containers, including two treatments with thirty replicates each, were processed in a laboratory scale mini-silo of 21 cm radius by 45 cm height. Three replicate containers were opened and sampled for analysis at 0, 0.5, 1, 2, 3, 4, 6, 10, 18 and 34 days after being ensiled. One silage container from each treatment was installed with a remote controlled electronic thermometer to record the temperature changes. Analysis included silage temperature, pH, fermentation acids, the water-soluble carbohydrates and chemical compositions and the silage protein fractions. Results showed that on the first day, the temperature of the ensiled corn was slightly higher than room temperature, but returned to room temperature on the second day. The pH and concentrations of WSC, ADF, lignin and acetic acid in the AFE treated silage were significantly lower than the control groups (p<0.05). The lactic acid and crude protein on the other hand were significantly higher in the AFE treated silage as compared to the control (p<0.05) at the end of the ensilage period. The DM content was significantly higher (p<0.05) whereas the butyric acid content of the AFE treated silage was significantly lower (p<0.05) than the control at the end of the 34 day ensilage period. Titratable acid and buffering capacity in the corn silage were not significantly different between treatment groups (p>0.05). Ammonia N concentration in the AFE treated silage showed a trend of decrease (p>0.05). NPN and the protein fraction A in both groups increased during the conservation period, but fraction A in the AFE treated corn silage was significantly higher than the control silage (p<0.05). During the conservation period, the AFE treated corn silage showed a trend toward a decrease in fractions $B_1$, $B_3$ and C (p<0.05). The protein fraction B2 showed a trend toward increase in the control group and an inconsistent trend in the AFE treated silage during the ensiling period. The AFE treated silage showed a better Flieg score over the control silage (97 vs. 75) as calculated from the concentrations of lactic acid, acetic acid and butyric acid.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.10
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• pp.1676-1684
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• 2004

Gel properties of recovered protein from mackerel, frozen blackspotted croaker, chicken breast and pork leg using acidic and alkaline processing were evaluated. Myofibrillar protein from mackerel by acidic processing did not form a heat-induced gel. However, the recovered protein including sarcoplasmic protein formed heatinduced gel. Breaking force of gel from mackerel processed at pH 10.5 was the lowest. A deformation value of frozen blackspotted croaker was the highest, followed by chicken breast, pork leg and mackerel. Whiteness of frozen blackspotted croaker was the highest among heat-induced gel. Breaking force, deformation and whiteness were decreased by addition of recovered protein from mackerel, but price was increased. A breaking force and whiteness of heat-induced gel added recovered protein from chicken breast were increased, and the price was greatly decreased. When the constraint of breaking force, deformation and price of raw material were set up above 110 g, 4.5 mm and below 2,000 won/kg. A optimum formulation for blending protein was 36∼50% for frozen blackspotted croaker, 34∼40% for chicken breast, 14∼25% for pork leg. The heat-induced gel of recovered protein from frozen blackspotted croaker showed compact structure compared to that of recovered protein from mackerel. A formulation of chicken breast and pork leg based on blackspotted croaker can be used in surimi based seafood products having various texture.

• Nutritional Sciences
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• v.8 no.1
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• pp.28-34
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• 2005

To examine the combined effects of a high-protein diet and aerobic exercise on body weight and composition and blood lipid profiles in overweight women, 30 young women were recruited and placed into three groups: The high-protein diet and exercise group (HPE), the exercise-only group (EXO) and the control group (CON) (30$\pm$3%, 27$\pm$2%, and 29$\pm$3% body fat, respectively) for an 8-week experimental period. Daily diet included 25% isolated soybean protein (>90% protein, approximately 400 kcal) combined with each subject s usual diet for the HPE group. The exercise program consisted of aerobic-type exercises undertaken >3 times/wk and for>30 min/session at 50-60% of maximal capacity. Physical fitness, body composition, serum total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), triglycerides (TG) and glucose were measured before and after the experiment. Maximal aerobic capacity increased by the end of experiment in both the HPE (from 27.2$\pm$3.5 to 35.l$\pm$5.9 ml/kg/min, p<0.01) and EXO (from 30.3$\pm$5.4 to 33.8$\pm$3.8 mㅣ/kg/min, p<0.05) groups. Percent body fat decreased by 3.3% (p<0.01) in the HPE group and by 1.5% (p<0.05) in the EXO group by the end of the experiment, but not in the CON group. Lower back strength and agility increased only in the HPE group. In the HPE group, TC decreased from 168$\pm$20 to 155$\pm$18 mg/dL and HDL-C increased from 57$\pm$l0 to 61$\pm$9 mg/dL in HPE (p<0.01). But TC and HDL-C did not change in the EXO and CON groups. TG and glucose did not vary among the groups. Although the EXO group showed a similar outcome to that of the HPE group, a favorable change in body composition and blood lipids as well as an improvement in aerobic capacity was more marginal in the latter group.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.3
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• pp.212-221
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• 2019

Sestrin-2 (SESN2) as a stress-metabolic protein is known for its anti-oxidative effects as a downstream factor of PERK pathways in mammalian cells. However, the expression patterns of SESN2 in conjunction with the UPR signaling against to ER stress on porcine oocyte maturation in vitro, have not been reported. Therefore, we confirmed the expression pattern of SESN2 protein, for which to examine the relationship between PERK signaling and SESN2 in porcine oocyte during IVM. We investigated the SESN2 expression patterns using Western blot analysis in denuded oocytes (DOs), cumulus cells (CCs), and cumulus-oocyte complexes (COCs) at 22 and 44 h of IVM. As expected, the SESN2 protein level significantly increased (p < 0.01) in porcine COCs during 44 h of IVM. We investigated the meiotic maturation after applying ER stress inhibitor in various concentration (50, 100 and 200 μM) of tauroursodeoxycholic acid (TUDCA). We confirmed significant increase (p < 0.05) of meiotic maturation rate in TUDCA 200 μM treated COCs for 44 h of IVM. Finally, we confirmed the protein level of SESN2 and meiotic maturation via regulating ER-stress by only tunicamycin (Tm), only TUDCA, and Tm + TUDCA treatment in porcine COCs. As a result, treatment of the TUDCA following Tm pre-treatment reduced SESN2 protein level in porcine COCs. In addition, SESN2 protein level significantly reduced in only TUDCA treated porcine COCs. Our results suggest that the SESN2 expression is related to the stress mediator response to ER stress through the PERK signaling pathways in porcine oocyte maturation.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.289-295
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• 2024

We have developed an aptamer that specifically binds to Porphyromonas gingivalis to reduce the cellular damage caused by P. gingivalis infection and applied it as a biosensor. P. gingivalis is one of the major pathogens causing destructive periodontal disease among the periodontal microorganisms constituting complex biofilms. Porphyromonas gingivalis G-protein (PGP) known to play an important role in the transmission of germs was used as a target protein for the screening of aptamer. The aptamer that has binds to the G-protein of P. gingivalis, was screened and developed through the Systemic Evolution of Ligands by Exponential Energy (SELEX) method. Modified-Western blot analysis was performed with the aptamer which consisted of 38 single-stranded DNA to confirm the selectivity. ELONA (enzyme linked oligonucleotide assay) used to confirm that the aptamer was sensitive to PGP even at low concentration of 1 ㎍/ml. For the rapid detection of P. gingivalis, we constructed a surface plasmon resonance biosensor with SPREETA using the PGP aptamer. It was confirmed that PGP could be detected as low concentration as at 0.1 pM, which is the minimum concentration of aptamer sensor within 5 min. Based on these results, we have constructed a SPREETA biosensor based on aptamer that can bind to P. gingivalis G-protein. It can be used as an infection diagnosis system to rapidly diagnose and analyze oral diseases caused by P. gingivalis.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.525-537
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• 2024

Pilins are protein subunits of pili. The pilins of type IV pili (T4P) in pathogenic bacteria are well characterized, but anything is known about the T4P proteins in acidophilic chemolithoautotrophic microorganisms such as the genus Acidithiobacillus. The interest in T4P of A. thiooxidans is because of their possible role in cell recruitment and bacterial aggregation on the surface of minerals during biooxidation of sulfide minerals. In this study we present a successful ad hoc methodology for the heterologous expression and purification of extracellular proteins such as the minor pilin PilV of the T4P of A. thiooxidans, a pilin exposed to extreme conditions of acidity and high oxidation-reduction potentials, and that interact with metal sulfides in an environment rich in dissolved minerals. Once obtained, the model structure of A. thiooxidans PilV revealed the core basic architecture of T4P pilins. Because of the acidophilic condition, we carried out in silico characterization of the protonation status of acidic and basic residues of PilV in order to calculate the ionization state at specific pH values and evaluated their pH stability. Further biophysical characterization was done using UV-visible and fluorescence spectroscopy and the results showed that PilV remains soluble and stable even after exposure to significant changes of pH. PilV has a unique amino acid composition that exhibits acid stability, with significant biotechnology implications such as biooxidation of sulfide minerals. The biophysics profiles of PilV open new paradigms about resilient proteins and stimulate the study of other pilins from extremophiles.

• Korean Journal of Food Science and Technology
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• v.6 no.2
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• pp.75-78
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• 1974

An experiment has been carried out in order to analyze the main components of Korean Cattles' milk, and fractionate the milk protein by DEAE-cellulose column. The results obtained were summarized as follow. 1) The average values of specific gravity, pH and acidity of Korean Cattles milk which were negative in alcohol test were 1,036, 6.4 and 0.21, respectively. 2) The average values of total solids, solids-not-fat, protein, lactose and ash contents of Korean Cattles milk were 11.61%, 9.53%, 2.08%, 3.99%, 4.76% and 0.86%, respectively. 3) Distribution of casein, whey protein, N.P.N., protein precipitated in 12% TCA, lactoglobulin and lactalbumin contents of the milk were 3.07%, 1.13%, 0.10%, 4.06%, 0.34% and 0.66%, respectively. 4) Acid casein obtained from Korean Cattles milk was fractionated into four fractions on DEAE-cellulose column with 0.005M tris-citrate buffer containing 6M urea, pH 8.6, and the ratio of the fraction I, II, III and IV was 3.24%, 52.67%, 26.22% and 17.87%, respectively. 5) Whey protein obtained from Korean Cattles milk was also fractionated into four fractions on DEAE-cellulose column with 0.04M phosphate buffer, pH 5.8, and the ratio of the fraction I, II, III and IV was 41.74%, 10.17%, 1.50% and 46.59%, respectively.

• The Korean Journal of Pharmacology
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• v.32 no.2
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• pp.243-257
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• 1996

CCK and cholinergic agonist stimulate enzyme release from the pancreatic acini via G-protein-mediated activation of phospholipase C, In contrast secretin and related peptides increase the level of cAMP and activate cAMP-dependent protein kinase. Camostat, a synthetic protease inhibitor, causes pancreatic hypertrophy and hyperplasia by increasing the CCK release. In this study, the secretagogue-induced changes of intracellular proteins were examined in the dispersed pancreatic acini of rats with or without camostat treatment. Camostat(FOY-305, 200 mg/kg, p.o.) was given for 4 days twice daily and the dispersed acini were prepared at 12 bouts after last treatment. The profiles of Intracellular phosphoproteins were analyzed by two-dimensional gel electrophoresis after incubating the acini with $^{32}P$. The amylase release from the dispersed acini was measured. The pancreatic weight was increased to 126% of control, while amylase activity per mg acinar protein decreased to 41% of control, The maximum response of amylase release from dispersed acini to CCK-8 or carbachol was markedly decreased(65% or 46% of control, respectively). The group of intracellular proteins(24 kD, pI $4.5{\sim}8.5$) was increased in quantity by camostat. CCK-8 or secretin increased phosphorylation of a protein(34 kD, pI 4.7) in camostat-treated as well as control rats. CCK-8 increased tyrosine phosphoryiation in the acini of control rats. However, in camostat-treated rats, the basal level of tyrosine phosphorylation was increased and it was rather decreased by CCK-8. Secretin had no effect on the level of tyrosine phosphorylation in acini. These results indicate that both phospholipase C and adenylate cyclase induce phosphorylation of an intracellular acinar protein(34 kD, pI 4.7) and camostat treatment increases the basal level of tyrosine phosphorylation in acinar cells. And these results suggest that not only serine/threonine protein kinase but also protein tyrosine kinase/phosphatase are involved in the process of CCK receptor mediated stimulation-secrelion coupling.