• Parasites, Hosts and Diseases
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• v.30 no.2
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• pp.133-140
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• 1992

Theileria sergenti were isolated from infected erythrocytes by hypotonic Iysis, and soluble meroBoite antigens were purified by sonication and differential centrifugation. The preparation contained 29, 34, 35 and 105 kD immuno-dominant polypeptides. The soluble antigens (0.5 mg/ml) were prepared and fortified with Freund's adjuvant. Five month old naive Korean calves were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later Nine weeks after the booster dose, vaccinates and controls were challenged with a homologous stabilate (5.6×106 RBC/dose, 40% Parasitemia). All animals were monitored for hematocrit, total erythrocyte count, parasitemia and for the specific antibody by Western immuno- blot (WB) and indirect immuno-auorescent antibody(IFA) test. By 18 weeks after vaccination (6 weeks after the challenge), vaccinated cattle had an average IFA titer of 1 : 10,240 compared with 1 : 1,280 of the controls. The vaccinates showed ne91igib1e change in hematocrit and total RBC count whereas control animals showed significant (P<0.05) hematological chanties and associated anemia. After vaccination and challenge, the antibody responses demonstrated that vaccination had induced significant production of antibody to the 29 and 35 kD polypeptides. The latter polypeptide was much more strongly recognized by the vaccinated animals, and thus it may be a potential candidate for the vaccine.

• Korean Journal of Weed Science
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• v.12 no.4
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• pp.368-373
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• 1992

The effect of alachlor treatment on protein synthesis was studied. Protein synthesis was inhibited by $1{\times}10^{-4}$ M and $1{\times}10^{-3}$M of alachlor 5.8% and 86.5%, respectively, while did not occur blow $1{\times}10^{-5}$M alachlor. Soluble protein of alachlor treated oat root tips was examined by polyacrylamide gel electrophoresis. The proteins extracted from oat root tips showed that they were made up of subunits blow 100 kd polypeptides by SDS-PAGE. As compared to control, high molecular proteins(above 47 kd) were inhibited of oat root treated with alachlor, while low molecular proteins(below 23 kd) were increased. Two-D gels showed that alachlor caused decrease(1-6 spots) or increase(7-10 spots) in number of polypeptides on silver staining. The intensity of some polypeptides of soluble proteins (molecular mass of 83 kd : 1, 2 spots, 70 kd : 3, 4 spots, and 47.5 kd : 5, 6 spots) decreased in alachlor treatment, whereas the intensity of other peptide bands (20 kd : 7 spot and 16 kd : 8, 9, 10 spots) increased. Oat root tip proteins present in the neutral zone are masked by diffusing of major proteins, but proteins in acid zone are resolved minor proteins.

• The Korean Journal of Food And Nutrition
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• v.3 no.2
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• pp.123-132
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• 1990

Sweet potato $\beta$-amylase is a tetrameric enzyme consisting of four identical polypeptide chains with a molecular weight of 5.6$\times$104, though most of the other $\beta$-amylases are monomeric enzymes. But, the relationship between subunit structure and catalytic function of the enzyme is not known. This study was done to know what the function of the subunit structure of the enzyme is. We obtained the monomer from the enzyme by the treatment of SDS, alkali pH buffer and urea. But the monomer had not activity. We tried to prepare the active monomer from the enzyme by the modification with periodate-oxidized soluble starch , In the result, we succeeded in isolating an active monomer as an oxidized soluble starch-conjugated form The active monomer had 57% of the original activity, 13.2% of the sugar and the molecular weight was estimated to be 5.4$\times$104. This results suggest that the tetrameric form of the enzyme is a most stable one and exists in nature, and the subunit structure of the enzyme Plays an important role in stabilization but not catalytic function.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.41 no.5
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• pp.563-568
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• 1996

This experiment was conducted to obtain basic information of biochemical changes in the process of senescence by measuring the total RNA, protein, protease activity and electrophoretic pattern of protein in tobacco plant. The content of soluble protein increased by 15 days after leaf emergence and its level was not changed from 15 to 35 days after leaf emergence. The content of total RNA showed a maximum value at 15 days after leaf emergence and then decreased rapidly until 30 days after leaf emergence. The activity of protease of neutral fraction was higher than that of acidic fraction and rapidly increased up to the end of senescence after 50 days after leaf emergence. According to the analysis of electrophoresis, polypeptide band of 61kd was developed after 35 days after leaf emergence and increased by the end of senescence.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.6
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• pp.564-570
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• 1994

This experiment was conducted to determine seed storage protein pattern and structural character of differed peanut cultivars during germination. Soluble protein content in both Namdae and Daekwang cultivars remarkably decreased in cotyledon site at 2 or 3 days after incubation(DAI) and in embryonic axis site at 1 or 2 DAI, showing 28∼29% in cotyledon site and 10% in embryonic axis site at 5 DAI. Protein subunits such as 66, 43, 40 and 35.5kD bands in the cotyledon site of Namdae and Daekwang cultivars disappeared, but 21.5-23kD band disappeared slightly, but low polypeptide band such as 14-16kD increased gradually, and the same trend has been obserbed in embryonic axis site during 2 DAI. The amount of new protein formed during germination period was highest in cotyledon site at 3 DAI, and in embryonic axis site at 2 DAI. 16kD bend detected in cotyledon site of Daekwang cultivar during germination.

• Animal cells and systems
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• v.15 no.1
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• pp.29-36
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• 2011

Insect lysozymes are basic, cationic proteins synthesized in fat body and hemocytes in response to bacterial infections and depolymerize the bacterial cell wall. The c-type lysozyme of the insect Spodoptera litura (SLLyz) is a single polypeptide chain of 121 residues with four disulfide bridges and 17 rare codons and is approximately 15 kDa. The full-length SLLyz cDNA is 1039 bp long with a poly(A) tail, and contains an open reading frame of 426 bp long (including the termination codon), flanked by a 54 bp long 5' UTR and a 559 bp long 3' UTR. As a host for the production of high-level recombinant proteins, E. coli is used most commonly because of its low cost and short generation time. However, the soluble expression of heterologous proteins in E. coli is not trivial, especially for disulfide-bonded proteins. In order to prevent inclusion body formation, GST was selected as a fusion partner to enhance the solubility of recombinant protein, and fused to the amplified products encoding mature SLLyz. The expression vector pGEX-4T-1/rSLLyz was then transformed into E. coli BL21(DE3)pLysS for soluble expression of rSLLyz, and the soluble fusion protein was purified successfully. Inhibition zone assay demonstrated that rSLLyz showed antibacterial activity against B. megaterium. These results demonstrate that the GST fusion expression system in E. coli described in this study is efficient and inexpensive in producing a disulfide-bonded rSLLyz in soluble, active form, and suggest that the insect lysozyme is an interesting system for future structural and functional studies.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.119-126
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• 1986

A periplasmic endoprotease, named protease Pi, was purified to homogeneity from Escherkchia coli by conventional procedure with insulin as substrate. This enzyme degrades insulin and glucagon to trichloroacetic acid-soluble meterials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. Its molecular weight was 110, 000 when determined by gel filtration on Sephacryl S-300 and was 105, 000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, it appears to be single polypeptide. This snzyme is metalloprotease, since it is completely inhibited by o-phenanthroline and can be activated by addition of divalent metal cations, such as $Mg^{2+}\;and\;Co^{2+}$. It is destinct from protease Ci, a cytoplasmic insulin degrading enzyme, since protease Pi is localized to the periplasm. Since protease Pi selectively degrades GTP cyclohydrolase I, it appears to play a role in the regulation of pteridine biosynthesis.

• The Journal of Natural Sciences
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• v.4
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• pp.1-10
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• 1991

The expression of the cloned Saccharomyces cerevisiae antioxidant protein gene in E. coil on plasmid was studied. Introduction of the DNA encoding yeast thiol-specific antioxidant protein into expression vector, PKK 223-3, having the tac promoter resulted in the production in E. coil of a Mr 27,000 polypeptide. This protein represented as much as about 1% of the bacterial soluble protein and showed the thiol-specific antioxidant properties. The bacterial protein showed physico-chemical properties indistringuishable from those of the yeast protein.

• Applied Biological Chemistry
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• v.18 no.4
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• pp.194-202
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• 1975

This study was conducted to study the properties of the water-soluble natural chelating agents from garbage compost and activated sewage sludge responsible for Fe chelation, which is closely associated with the effectiveness in correcting iron chlorosis in plant. The water-soluble fraction of these materials was fractionated by menas of Sephadex gel filtration and the fractions of Fe chehates were traced by radioactive $^{59}Fe$. The fractions were examined by ultraviolet and infrared. spectroscopy and stability constants for Fe. The water-soluble fraction from garbage compost was separated by Sephadex G-25 into approximately four fractions. Most of the added $^{59}Fe$ was associated with fraction I, which appeared at the void volume. Further fractionation by Sephadex G-50 indicated that the molecular weight of water-soluble chelating agents is in the approximate range of 5000 to 10,000. The water-soluble fraction from activated sewage sludge gave six fractions by Sephadex G-25. Most of the added $^{59}Fe$ was found in the fraction I,II, and III, The molecular weights of most chelating agents associated with $^{59}Fe$ appeared to be less than 5,000 and those of fraction I that appeared at the void volume was in the range of 5,000 to 1,000. Discrepancy between radio activity count and UV absorption indicated the heterogeneity of the fractions obtained by Sephadex gel filtration. Ultraviolet absorption spectra of all fractions separated by Sephadex G-25 and containing chelating agents showed no differences. Fraction IV and V of sewage extract showed absorption maxima and shifting similar to nucleic acid components suggesting the presence of decomposition products of nucleic acid. Similarity fraction VI contained phenolic type amino acid groups. Fraction I of compost extract contained most of the added $^{59}Fe$ and showed weak but extra definite absorption in the 1230, and $1270cm^{-1}$ region, suggesting that extra oxygen groups in polyphenolic structure were probably involved in Fe chelation. In sewage extract, fraction I,II, and III in which most of the $^{59}Fe$ was found, showed strong definite polypeptide absorption in the region of $1540cm^{-1}$ due to NH deformation and C-N stretching of amide groups in the peptidebond. These extra functional groups in fraction I, II, and III appeared to be associated with Fe chelation. The other fractions, not associated with $^{59}Fe$, still have carboxyl and hydroxyl groups, suggesting that these functional groups in these water extracts may not independently form the Fe chelates. Precipitation of ferric hydroxide precluded measuring the stability constants for Fe-chelates. However, the formation constants for Zn chelates as log K values for compost extract and sewage extract at pH 4.0 from which the strength of chelation with Fe could be presumed, were 8.23, and 9.75, respectively, indicating strong complexation with metals. The chelating capacity of compost extract containing 6.5 g organic matter per liter was 0.82 mM, and that of sewage extract containing 5.3 g per liter was 0. 64 mM.

• Korean Journal of Environmental Agriculture
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• v.17 no.4
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• pp.296-300
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• 1998

The analysis of biochemical changes in tobacco plant as increase of NaCl concentraion was conducted. Total protein content and soluble protein content were decreased as NaCl concentration was increased, in that steady decreased until 120mM NaCl and largely decreased at 150mM NaCl. The expression of 74Kd subunit was increased until 60mM NaCl. However, the amount of 74Kd protein was decreased from 90mM NaCl. There was no difference for expression of other protein subunits. Chlorophyll a content was significantly decrease as NaCl concentration was increased, but chlorophyll b content was not much decreased. The slow increase up to 120mM NaCl and large increase at 150mM NaCl for ATPase and peroxidase activities indicated that 120mM NaCl could be a limiting concentration for salt injury.