• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.18 no.12
• /
• pp.575-580
• /
• 2017

This study analyzed the effects of RGG box of hnRNP A1 on its subcellular localization and stabilization of hnRNP A1 over a three year period from October 2014. First, a 6R/K mutation in RGG box was generated, and pcDNA1-HA-hnRNP A1(6R/K) was constructed. The subcellular localization of hnRNP A1(6R/K) from the HeLa cells transfected with this plasmid DNA was analyzed by immunofluorescence microscopy. HA-hnRNP A1(6R/K) was found to exhibit nuclear and cytoplasmic fluorescence. The stability of hnRNP A1(6R/K) was checked by Western blot analysis using the expressed protein from the HeLa cells transfected with the pcDNA1-HA-hnRNP A1(6R/K). The results show that HA-hnRNP A1(6R/K) has a smaller size. These confirm that HA-hnRNP A1(6R/K) is localized both in the nuclear and cytoplasm, not because 6R/K mutation affects the nuclear localization of hnRNP A1, but because 6R/K mutation causes hnRNP A1(6R/K) to cleave at the mutation or near the mutation site. The cleaved protein fragment, which lacks the M9 domain (i.e. nuclear localization signal of hnRNP A1), did not exhibit nuclear fluorescence. This suggests that the arginines of RGG box in hnRNP A1 play an important role in stabilizing hnRNP A1. An analysis of the RNA-binding ability of hnRNP A1(6R/K) expressed and purified from bacteria will be a subsequent research project.

• Reproductive and Developmental Biology
• /
• v.32 no.3
• /
• pp.153-158
• /
• 2008

Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 1995.06b
• /
• pp.77-96
• /
• 1995

To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

• Food Science and Preservation
• /
• v.18 no.5
• /
• pp.729-738
• /
• 2011

This study was conducted to investigate the effect of extruded pellets (EP) containing different levels of protein (51%-55%) and lipid (9%-15%) on flesh quality of flounder (Paralichthys olivaceus) comparing with raw fish-based moist pellet (MP). Two replicate groups of 40 fish per each tank (initial mean weight 106 g) were fed one of three experimental EP (EP1, EP2 and EP3) containing different protein and lipid levels, a commercial EP (EP4) and MP for 16 weeks. Moisture content of fish fed MP was significantly higher than that of fish fed all EP. Significantly higher contents of 16:1n-7, 20:4n-6, 20:5n-3 and lower 18:2n-6 were observed in fish fed EP4 and MP compared with fish fed EP1, EP2, EP3. The 22:6n-3 content was not significantly different among all groups. Taurine content of fish fed MP was significantly higher than that of fish fed all EP. Asparagine content of fish fed EP1 was significantly higher than that of fish fed EP2, EP, EP4 and MP. Significant difference were observed in lysine and serine contents of fish fed experimental diets. In nucleotides and their related compounds, ATP and AMP content was not affected by diets. Significant difference were observed in IMP contents of fish fed experimental diets. Textural properties, no significant difference was observed among the fish groups fed different diets.

• Korean Journal of Food Science and Technology
• /
• v.29 no.3
• /
• pp.452-459
• /
• 1997

Effect of extraction pH on mechanical properties such as tensile strength (TS) and elongation (E) and on water vapor permeability (WVP) of soybean protein isolate (SPI) edible films extracted from soybean meal was investigated. Five pHs, acidic range (pH 2.0 and pH 3.0), neutral range (pH 7.0) and alkalic range (pH 10.0 and 12.0), were used to extract SPI. TS of the film extracted at pH 7.0 was the lowest, and WVP of $SPI_3$ (SPI extracted at pH 3) film was the lowest value among the films. The WVP of $SPI_3$ films was $3.349\;{\times}\;10^{-10}\;g{\cdot}m/m^2{\cdot}s{\cdot}Pa$ and increased to $3.871\;{\times}\;10^{-10}\;g{\cdot}m/m^2{\cdot}s{\cdot}Pa$ as film thickness increased from $55\;{\mu}m$ to $72\;{\mu}m$ thickness. Three different plasticizers (glycerol, polyethylene glycol and propylene glycol) were used for $SPI_2$ (SPI extracted at pH 2) film. TS of $SPI_2$ films was 12.297 MPa and decreased to 1.356 MPa for glycerol and showed the same trend in other two plasticizers. The SPI films extracted at acidic range were shown higher mechanical properties and lower water vapor permeabilities than those of extracted at neutral and alkalic ranges. The difference of SPI film properties seemed to be attributed by 11S/7S ratio as well as protein content.

• Journal of Nutrition and Health
• /
• v.7 no.2
• /
• pp.53-63
• /
• 1974

For the elucidation of the nutritional status of residences of apartments located in various areas in Seoul city, the food and nutrition survey was undertaken by the graduate students from Department of Food & Nutrition, Ewha Womans University in 1973. The socioeconomic stratification was mainly considered for the selection of sample apartments included in this study. Hangang Mansion, KIST, Munwhachon & Bukahyun apts were selected in this respect. A. Common living circumstances. Through the questionaries, author concluded nutritional concept on everyday food life as follows; 1. Higher income seems less effective in everyday food life on the nutritional view point than higher education does. 2. It needs to be urgently improved to be accustomed to use exact measuring concept of foods when they cook. 3. It needs to be improved to serve meals for individual portion at meal table, because the portion control at meal table will effect to national food policy. B. Nutrition Survey. Average daily calroie intake was the highest in Hangang Mansion and lowest in Bukahyun apt., in accordance with their economic living standard, but most of their calorie intakes were lower than the R.D.A. (3000ca1 for male, 2200 Cal for female). But the composition ratio of the total calorie intake was near to the ideal, carbohydrate 60%, protein 15%, fat 25%. Although there were differences in their economic living situations in urban area, most people ingested relatively good quality and large amounts of protein and fat. Vitamin A and Niacine intake exceeded the R.D.A., but the intake of Ca, Thiamine, Riboflavin, Ascorbic Acid were far less than that of R.D.A. C. food Intake. Amounts of total daily food intake were highest in Bukahyun apt. and lowest in Hangang Mansion. These showed adverse aspects to the total calorie intakes and their income levels. The ratio of cereals, $40%{\sim}50%$ of the total food intake, was higher than any other kinds of foods in all apartments. Among the other foods except cereals, there represented the highest level of meats in Hangang Mansion and KIST apt., while intake of Kimchies were highest in Munwhachon and Bukahyun apt. In the case of vegetables and fruits intake, the ratio of them in Hangang Mansion and KIST apt. were high, on the other hand the ratio of starchy roots appeared lowest in Hangang Mansion. For the main sources of protein, meats were ingested in Hangang Mansion and KIST apt., meats and fishery were used in Munwhachon apt., and beans were eaten highly in their meals in Bukahyun apt. These food contents and distributions showed the significant differences not only their economic classes but also regional characteristics.

• Journal of Life Science
• /
• v.19 no.11
• /
• pp.1529-1537
• /
• 2009

To elucidate further the antitumor effects of a natural L-arginine analogue, L-canavanine, the mechanism underlying apoptogenic activity of L-canavanine and its modulation by protein tyrosine kinase $p56^{lck}$ was investigated in human Jurkat T cells. When the cells were treated with 1.25 to 2.5 mM L-canavanine for 36 h, several apoptotic events including mitochondrial membrane potential (${\Delta\Psi}m$) loss, activation of caspase-9, -3, -8, and -7, poly (ADP-ribose) polymerase (PARP) degradation, and DNA fragmentation were induced without alteration in the levels of Fas or FasL. These apoptotic changes were more significant in $p56^{lck}$-deficient Jurkat clone JCaM1.6 than in $p56^{lck}$-positive Jurkat clone E6.1. The L-canavanine-induced apoptosis observed in $p56^{lck}$-deficient JCaM1.6 cells was significantly reduced by introducing $p56^{lck}$ gene into JCaM1.6 cells by stable transfection. Treatment of JCaM1.6/lck cells with L-canavanine caused a transient 1.6-fold increase in the kinase activity of $p56^{lck}$. Both FADD-positive wild-type Jurkat T cell clone A3 and FADD-deficient Jurkat T cell clone I2.1 exhibited a similar susceptibility to the cytotoxicity of L-canavanine, excluding involvement of Fas/FasL system in triggering L-canavanine-induced apoptosis. The L-canavanine-induced apoptotic sub-$G_1$ peak and activation of caspase-3, -8, and -7 were abrogated by pan-caspase inhibitor (z-VAD-fmk), whereas L-canavanine-induced activation of caspase-9 was not affected. These results demonstrated that L-canavanine caused apoptosis of Jurkat T cells via the loss of ${\Delta\Psi}m$, and the activation of caspase-9, -3, -8, and -7, leading to PARP degradation, and that the $p56^{lck}$ kinase attenuated the ${\Delta\Psi}m$ loss and activation of caspases, and thus contributed as a negative regulator to L-canavanine-induced apoptosis.

• Tuberculosis and Respiratory Diseases
• /
• v.42 no.6
• /
• pp.855-861
• /
• 1995

Background: Parathyroid hormone-related protein(PTHrp) was first identified as the cause of hypercalcemia in malignancy. Hypercalcemia can be found in malignancy, especially in the epidermoid carcinoma of the lung, even without extensive metastases to the bones. The application of sensitive assays for PTHrp may help in the early diagnosis of lung cancer, in the monitoring of treatment and in the detection of recurrence. Method: Serum PTHrp was measured by radioimmunoassay detecting the N-terminal 1~34 peptide of human PTHrp(PTHrp 1-34) in 63 histologically confirmed lung cancer patients and 22 healthy controls. Result: Serum PTHrp(mean$\pm$S.E.) was $312{\pm}68.9pg/ml$ in 63 lung cancer patients and $158{\pm}38.2pg/ml$ in 22 controls(p>0.05). PTHrp was $356{\pm}103.9pg/ml$ in 34 epidermoid carcinoma patients, $281{\pm}148.7pg/ml$ in 15 adenocarcinoma patients and $316{\pm}140.8pg/ml$ in 9 small cell carcinoma patients. In epidermoid carcinoma patients, PTHrp was $570{\pm}472.3pg/ml$ in stage II(n=3; p<0.05 vs controls), $166{\pm}22.4pg/ml$ in stage IIIa(n=9), $282{\pm}113.3pg/ml$ in stage IIIb(n=12) and $668{\pm}367.9pg/ml$ in stage IV(n=9; p<0.05 vs controls). PTHrp was significantly increased in 8 epidermoid carcinoma patients with bone metastases($1526{\pm}811.2\;pg/ml$; p<0.0005 vs controls). Hypercalcemia was observed in an epidermoid carcinoma patient whose PTHrp value was 244 pg/ml. Conclusion: The serum PTHrp was increased in advanced epidermoid carcinoma patients even without hypercalcemia. The measurement of PTHrp may be not helpful in the early diagnosis of lung cancer. But the lung cancer should be suspected in the marked elevation of PTHrp. It may be of value in detecting patients of advanced diseases with bone metastases or patients who might develop the malignancy associated hypercalcemia.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.12 no.2
• /
• pp.122-136
• /
• 1983

The effect of dietary casein, calcium, magnesium and some vegetable oils such as seasme, perilla and soybean oil on the serum cholesterol level in the rabbit were studied for a period of 5 weeks using isocalories and isonitrogenous as basal diets. The experimental rabbits fed the following basal diets containing crude protein 68.47%, carbohydrates 13.35%. fats 16.18% and vegetable oil 10%. casein 10%, calcium and magnesium according to experimental plan making. In order to calculate the feeding efficiency, protein efficiency and calorie efficiency during period, the body weight gains were measured at the same time using same balance, respectively. The results are summarized as follows. Body weight gains per week of the group fed perilla oil, calcium and basal diet were the higher than any other groups. And body weights gains per week of the group fed basal diet, vegetable oil were the lower than any other groups. In the case of efficiency of reed, protein and calorie, the efficiency ratios of the group fed perilla oil were the higher than any other groups. Especially, perilla oil and calcium diet effect on body weight gain in rabbit. In the case of serum protein, the total proteins in serum were almost same value for all the groups. Serum albumin of group fed basal diet. vegetable oil and casein were the higher than any other groups. The ${\alpha}$-globulin of the groups, fed basal diet and calcium was the lower than any other grosps. The ${\beta}$-globulins of the groups fed basad diet, perilla oil and casein were the highest value. In serum lipoprotein, lipalbumin was almost same value for the groups fed vegetable oil, but fed vegetable oil and calcium diet was the lowest value. The ${\beta}$-lipoprotein in high cholesterol level group was increased some degree, the group fed perilla oil added was lower. The ratio of ${\beta}$-lipoprotein per lipalbumin was from 0.11 to 0.26. The ratio of lipalbumin per total lipoprotein was high in calcium and soybean oil added diet. In serum triglyceride, the level of triglyceride of groups fed seasme oil or perilla oil was the higher than any othe groups, but in the group fed casein and calcium or magnesium, the level of triglyceride level was decreased. Calcium and magnesium effect on triglyceridge level lowering action. In serum total cholesterol, the group does fed vegetable oil with basal diet and casein added more, total cholesterol level increased as much as triglyceride level increased. But the group does fed perilla oil and magnesium shows total cholesterol level decreased remarkably. In the group fed basal diet with calium, the amount of serum calcium was increased, but of serum magnesium was decreased. In the case of blood glucose, the group fed basal diet and vegetable oil was decreased. According to the regression and correlation coefficient in blood components in rabbit, there are positive correlation $${\gamma}{\sim_=}1$$ between serum cholesterol and triglyceride, ${\beta}$-lipoprotein, ${\alpha}$-globulin, calcium and magnesium according to diet composition. From the above results, the serum cholesterol level lowering factors in rabbit, was the amount of triglyceride and ${\beta}$-lipoprotein which was decreased in perilla oil fed. It assumes that serum cholesterol and triglyceride level lowering factors are not only unsaturation degree of fatty acid but the amount of calcium and magnesium and the ratio calcium per magnesium 2:1 in the diet.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.35 no.1
• /
• pp.68-77
• /
• 2006

This study was peformed to document the association between nutrient intakes, body mass index (BMI), waist/hip ratio (WHR), and a major risk factor for chronic diseases. A three-day dietary intake survey, using a 24 hour recall method, was obtained from 187 subjects aged 46 to 84 (mean age 65.3) living in Wando island area. The average daily mean energy intakes were 1869.0 kcal for male and 1943.9 kcal for female, respectively. Daily intakes of protein for male and female were 28.0 and 30.4 g, and those of fat were 31.5 and 28.51 g, respectively Carbohydrate dependency was decreased with age. Protein dependency was increased with age. The mean intakes of energy, protein, Vit. A, Vit. D, Vit. E, Ca, Zn did not meet Korean RDA for elderly. The level of serum triglyceride was higher in males than in females and showed the tendency to increase with age in both sexes, whereas HDL-cholesterol tended to decrease with age in both sexes. The levels of serum total-cholesterol and LDL-cholesterol were significantly higher in males than in females, particularly in the age of $46\~59$ (p<0.05). The level of atherogenic index (AI) was significantly higher in females than in males, particularly in the age of 80 and over (p<0.05) Based on these results, it is evident that people in island area did not consume enough nutrient. Specially, dietary intake of protein was not adequate. This study implies that triglyceride, total-cholesterol, LDL-cholesterol, AI were increased with increasing age, BMI and WHR.