• Journal of Ecology and Environment
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• v.41 no.9
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• pp.265-270
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• 2017

Background: Over the years, pine pollens have been excluded as an allergen due to its relatively large size, low protein content, and waxy hydrophobic layer, despite their abundance. However, recent studies suggest the possibilities of pine pollens being allergens, and it has been reported that allergy symptoms were highly prevalent in areas with considerably large pine forests and high possibility of exposure to the pollen. Therefore, we conducted a comparative analysis of the allergenicities of the pollens from the dominant species of Korean pines, red pine (Pinus densiflora), black pine (Pinus thunbergii), and pitch pine (Pinus rigida), in mice. Methods: The protein composition of the pollens from the three pine species was compared via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The pine pollens and proteins extracted from the pollens were introduced to BALB/c mice by nasal inhalation and application to exposed skin and the IgE produced by the mice were extracted from blood and analyzed via ELISA. Results: SDS-PAGE showed differing protein compositions of the pollens of the three pine species. Analysis of blood IgE compositions showed a similar amount of IgE produced when pollens were applied to skin. In contrast, when mice inhaled the pollens, P. densiflora was shown to induce significantly more IgE production than those of the other two species. Conclusions: The experimental results demonstrate that the pollens of all three South Korean pine species induce IgE production, and this production was more pronounced when the pollens were inhaled than when they were applied to the skin. Of the three species, the pollen of P. densiflora was found to induce the highest level of IgE production.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.2
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• pp.285-291
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• 1999

To confirm the food quality of Urechis unicinctus which have been favored as a special raw seafood in southern area in Korea, the chemical composition of Urechis unicinctus and those glycoprotein were studied. Freeze dried Urechis unicinctus was composed of more than 70% of crude protein and 15% of total carbohydrate. The amino acids such as glycine(18.69%), glutamic acid(12.50%) and aspartic acid(9.37%) were noted as major components of total amino acids. The predominant free amino acids were alanine(32.98%), glycine(27.50%), asparagine(19.65%) and taurine(8.29%), and the sum of them were more than 88% to total free amino acids, so they may cause unique taste of Urechis unicinctus with sweetness. As the basis of sugar composition analysis, 56.35% of glucose and 30.49% of N acetylglucosamine were contained respectively, and they might also play an important role in a sweet taste. The leading carbohydrate moiety of glycoprotein from Urechis unicinctus was identified as glucose and N acetylglucosamine similarly to raw muscle, and they occupied more than 50% of total sugar content. Fucose(18.32%) and D glucuronic acid(12.31%) also detected in higher levels com pared to raw muscle. The total amino acid profile of glycoprotein showed a similar trend to raw muscle protein but glycine was detected a significantly lower than that in raw muscle. The glycoprotein from Urechis unicinctus was composed of 4 kinds of subunits, i.e. 25kDa, 20kDa, 18kDa and 12.5kDa of molecular weights through the SDS polyacrylamide gel electrophoresis. On the basis of the IR spectrum of absorptions appeared in 1,240cm-1 and 850cm-1, the glycoprotein had sulfate groups.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.4
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• pp.348-352
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• 1992

Investigation on the characteristics of actomyosin was prepared from leg and breast muscle of duck treated by hard scalding and subscalding method and their extractability , ATPase activity , solubility and SDS polyacrylamide gel electrophoresis were compared. The extractability of actomyosin in leg and breast muscle of duck by hard scalding was 7.84 and 39.84mg/g, whereas 4.79 and 28.04mg/g by subscalding respectively. Ca-ATPase activity of breast muscle wash higher than that of leg muscle. In case of leg muscle, hard scalding was higher tan subscalding. Breast muscle showed that subscalding was higher than hard scalding in less than ionic strength 0.08, and was lower than hard scalding in over ionic strength 0.08.Mg-ATPase was great in ionic strength and subcalding was relatively higher than hard scalding. Without regard to be treated method and part, the start point and end point of solubility were like. Hard scalded muscle and breast muscle showed that proteins in thin filament produced many extraction.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.3
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• pp.287-299
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• 1989

This research was conducted in order to investigate thermal stability and nutritional value of corn lipids and proteins during thermal processing. The lipids of raw and popped corn were fractionated and analyzed by column and gas chromatography. The effect of thermal processing on corn proteins was examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and amino acid analysis. There was no remarkable change in proximate compositions during thermal processing. The lipid fractions obtained by silicic acid column chromatography were composed of neutral lipid(93.5%), glycolipid(3.8%), and phospholipid(2.7%), Although the thermal processing showed the increase in the ratio of unsaturated/saturated fatty acid, there was no significant differences in the lipid composition between raw and popped corn. Most of each protein fractions had lower molecular weight than 25,000 dalton and albumin fractions were distributed in the molecular weight range 11,500-94,000 daltons. Popped corn proteins did not show marked differences in their electrophoretic migrations when compared with raw corn proteins.

• Korean journal of applied entomology
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• v.23 no.3 s.60
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• pp.137-141
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• 1984

Crude sap, which was extracted from six Amaranthaceae plants, inhibited local lesion formation on Nicotiana glutinosa by tobacco mosaic virus(TMV) infection. Especially the remark. able inhibitory effect to TMV infection was shown on leaves of N. glutinosa precoated with the sap from Amaranthus mangostanus. The inhibitory activity of the sap from A. mangostanus was stable to storage in vitro for I day and to dilution 1/4 of the sap with distilled water. However, its activity was lost when the sap was heated at $70^{\circ}C\;to\;100^{\circ}C$ for 10 minutes. When the leaves of N. glutinosa precoated with the sap were sprayed with water, the inhibitory effect to TMV infection was maintained for 2 days. The A. mangostanus sap readjusted pH 3, pH 5, or pH 9 with 1 N HCl or 1 N NaOH did not decline the inhibitory action but the sap absorbed with $5\%\;to\;15\%$ charcoal completely lost their action. The protein components purified from A. mangostanus sap revealed three major bands by $5\%\;to\;15\%$ polyacrylamide gel electrophoresis and the top component of which showed the inhibitory action to TMV infection.

• Korean journal of applied entomology
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• v.27 no.4
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• pp.211-218
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• 1988

Polyhedral proteins and the endogenous alkaline protease associated with larval-derived polyhedra of nuclear viruses isolated from Spodoptera litura, Bombyx mori, and Hyphantria cunea were investigated. Polyhedral proteins prepared under alkaline protease heat-inactivated condition were separated as one band with 31Kd in S. litura a H. cunea NpV and 30Kd in B. mori NPV by the SDS-polyacrylamide gel electroptoresis. Whereas polyhedral proteins without heat-inactivation were degraded into smaller polypeptides with a certain pattern in alkaline solution. The results of double-immunodiffusion and western blot analysis with antisera against polyhedral proteins indicated that those three polyhedral proteins had common antigenic determinants and the degradation of polyhedral proteins by alkaline protease could be confirmed.

• Korean journal of applied entomology
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• v.30 no.3
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• pp.180-186
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• 1991

This study was carried out to acquire some basic biochemical informations on the granulosis virus(GV) of Pieris rapae and Pieris brassicae. The capsule protein was composed of a single polypeptide with a molecular weight of 30,000 dalton for P. rapae GV and 31,000 dalton for P. brassicae GV. The major amino acids of capsule protein were glutamic acid, aspartic acid and lysine. When the capsule protein was partially digested with trypsin, chymotrypsin, papain or Staphylococcus aureus V8 protease, the digested products of the two viruses showed no difference in electrophoretic mobility. The patterns of the polypeptides of the two virus particle on SOS-polyacrylamide gel showed a little difference in high molecular weight region(over MW 100 kd).

• Interdisciplinary Bio Central
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• v.4 no.3
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• pp.8.1-8.7
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• 2012

The motor neuron degeneration 2 (mnd2) mice carry a point mutation of A to T nucleotide transversion at the serine 276 residue of high temperature requirement A2 (HtrA2), resulting in losses of an AluI restriction enzyme site (5'AGCT3') and the HtrA2 serine protease activity. Moreover, dysfunctions of HtrA2 are known to be intimately associated with the pathogenesis of neurodegenerative diseases, including Parkinson's disease. Thus, this mnd2 mouse is an invaluable model for understanding the physiological role of HtrA2 and its pathological role in neurodegenerative diseases. Nevertheless, many molecular and cellular biologists in this field have limited experience in working with mutant mouse models due to the necessity of acquired years of the special techniques and knowledges. Herein, using the mnd2 mouse model as an example, we describe easy-to-use standard protocols for web-based analyses of target genes, such as HtrA2, and a novel approach for simple and accurate PCR-AluI-RFLP-based genotype analysis of mnd2 mice. In addition, band resolution of AluI-RFLP fragments was improved in 12% polyacrylamide gel running in 1X Tris-Glycine SDS buffer. Our study indicates that this PCR-AluI-RFLP genotype analysis method can be easily applied by the molecular and cellular biologist to conduct biomedical science studies using the other mutant mouse models.

• Biomedical Science Letters
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• v.15 no.1
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• pp.37-45
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• 2009

Insect cell cultures have become important tools in the production of biological substances for use in a variety of research, human and veterinary medicine, and pest control applications. These applications often require the introduction of foreign DNA into the cells and have generally used methods originally developed for use with human and other mammalian cell cultures. While these methods can be successfully employed, they are often less efficient with insect cells and frequently involve complex procedures or require specialized equipment. Even when they do work, they may require substantial modification because of differences in the culture medium or growth patterns of insect cells. In this study, We have optimized transfection conditions of Sf9 cell line using insect expression vector pIZT/V5-His which expresses green fluorescent protein effectively. Human stem cell factor (hSCF) is a glycoprotein that plays a key role in hematopoiesis acting both as a positive and negative regulator, often in synergy with other cytokines. It also plays a key role in mast cell development, gametogenesis, and melanogenesis. It can exist in membrane-bound form and in proteolytically released soluble form. As determined by an enzyme-linked immunosorbent assay performed, hSCF level in supernatant averaged 995ng/ml. The human hSCF was partially purified by immunoaffinity chromatography and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The results show that the hSCF has N-linked carbohydrate and corresponds to the soluble form, at or about 223 amino acids in length. The findings suggest functional importance for soluble hSCF in cells.

• BMB Reports
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• v.29 no.6
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• pp.555-563
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• 1996

A membrane-bound phosphatidylinositol 4-kinase (PI 4-kinase) was separated in a sucrose gradient and solubilized with 1% Triton X-100 from mouse brain. The enzyme was purified 2,952-fold by various chromatographic techniques including DEAE-cellulose, PI-Sepharose and Sephacryl S-200 gel filtration. The molecular weight of PI 4-kinase was approximately 76 kDa by gel filtration and 70.8 kDa by SDS-polyacrylamide gel electrophoresis. The purified enzyme exhibited specific activity of 11.2 nmol/min/mg protein and pi value of 4.7. Kinetic analysis of the PI 4-kinase indicated apparent $K_m$, values of 190 ${\mu}M$ and 120 ${\mu}M$ for phosphatidylinositol and ATP, respectively. The maximal activity of this purified enzyme was observed at pH 7.4 at an incubation temperature of $37^{\circ}C$. The enzyme activity was significantly activated by $Mg^{2+}$, $Mn^{2+}$ and $Fe^{2+}$, and inhibited severely by $Ca^{2+}$. PI 4-kinase was proved to be pure in its immunoblot test by polyclonal antibody prepared from immunized rabbit sera. By this test, we were able to detect the existence of the same type of PI 4-kinase from other mouse organ tissues, such as liver, heart, kidney and spleen. Furthermore, similar immunoblot analysis with the same antisera recognized the different epitopes of PI 4-kinase proteins from various organs of rabbit, chinese hamster and rat.