• Korean Journal of Microbiology
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• v.53 no.2
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• pp.79-82
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• 2017

An inky cap, Coprinus comatus synthesizes and secretes a laccase in the liquid yeast extract peptone dextrose medium. We have successfully purified the enzyme through the ion-exchange chromatography and the preparative gel electrophoresis. The estimated molecular weight was 67 kDa by the SDS-PAGE analysis. Optimum pH was pH 4.3 and optimum temperature was $25^{\circ}C$. The Km value was 0.45 mM and the Vmax was 0.0132 OD/min/unit for o-tolidine. Purified laccase showed 49.3% decolorizing activity against remazol brilliant blue R (RBBR) and 41.6% decolorizing activity against Poly R-478 after 12 h incubation.

• Food Science of Animal Resources
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• v.42 no.1
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• pp.73-83
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• 2022

The aim of this study is to determine the effects of using emulsion manufactured with soybeans (ES) to substitute chicken breast in Vienna sausages. Four types of Vienna sausages (S1: 10% ES and 50% chicken, S2: 20% ES and 40% chicken, S3: 30% ES and 30% chicken, and S4: 40% ES and 20% chicken) for this study were made. The pH, color, proximate composition, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), microphotographs, cooking yields, and texture profile analysis of sausages were examined. The pH value of uncooked and cooked sausages increased significantly with increasing ES content (p<0.05). The crude protein contents of S2, S3, and S4 were significantly higher than that of the control (p<0.05). Furthermore, the SDS-PAGE results showed that α-conglycinin, β-conglycinin, and the acidic subunit of glycinin all increased with increasing ES content. Microphotographs revealed that increasing the ES content decreased the size of fat globules. The cooking yields of samples increased significantly with increasing ES content (p<0.05). The hardness values of ES treated samples were significantly lower than that of the control (p<0.05). Therefore, 30% substitute of chicken breast with ES can improve the quality and structure of Vienna sausage, without inducing critical defects.

• Journal of the Korean Society of Tobacco Science
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• v.23 no.2
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• pp.103-108
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• 2001

Hog cholera virus(HCV) was purified from virus infected Bovine kidney cells. From this virus, total protein was analyzed by SDS-PAGE gel electrophoresis and about 55 kDa band of E2 envelope protein was detected. The viral RNA was purified and E2 cDNA was amplified by RT-PCR. E2 cDNA fragment was cloned to PCRII-TOPO cloning vector and named pE2. The analysis of nucleotide sequence showed that this E2 cDNA fragment inserted into pE2 was 1191 nucleotides long and coded 397 amino acids.

• Korean Journal of Animal Reproduction
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• v.16 no.4
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• pp.289-299
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• 1993

The polypeptide patterns of cellular and follicular components were analysed by SDS-PAGE and two dimensional(2-D)electrophoresis combined with isoelectric focusing (IEF) to establish protein profiles in each of the components in porcine follicles. Oocyte-cumulus complexes were cultured in M16+FCS+Gn at 39 in an atmosphere of 5% CO$_2$, in air for 35 h. At the end of the culture, the zona-free oocyte, ZP alone and cumulus cells were prepared and analysed either on 10% SDS-PAGE for the protein profile at the first dimensional gel or 2-D protein pattern. The amounts of each samples were determined for the visualization with Coomasie brilliant blue (CBB) or silver staining, thus giving useful information for the identification of specific proteins in the components or appropriate amount of samples for proper visualization. Oocyte showed 25 and 114 kd major protein band. Other minor components were additionally visualized with CBB on the same gel after silver staining procedure. Cumulus cells also showed specific proteins which is not present in the oocytes. The number of cumulus cell was proper to give major bands with CBB and additional minor bands with silver staining. To establish the degree of contamination from the remnant of the corona radiata to the ZP, zonae were differently prepared or analysed by SDS-PAGE.The preparation of the ZP in this study did not showed any contamination judged by the protein profile of the components. Also follicular fluid showed its specific protein profile without any significant differences among the different sizes of follicles. The established protein profile of each follicular component should be helpful for the identification and elimination of contaminated components, i. e., antigen preparation or immunological studies. The results also suggest that the preparation of each components in the study was appropriate and can be used for a further sensitive biochemical analysis in mammalian oocytes and early embryos.

• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.1-8
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• 1989

Serodiagnosis of parasitic infections is widely used, since parasites or their eggs are not always detected by ordinary methods. The sensitive tests such as ELISA are highly dependent on the purity of antigens used. To solve this problem. many workers have tried to find species-specific components of antigens, The present study was performed to determine the antigenic profile of crude saline extracts of 3, 5, 8 and 12-week old p. westermani worms, which were collected from experimentally infected cats, based on SDS-PAGE and immunoblot technique. The results were as follows: 1. The SDS-PAGE showed at least 30 Protein bands ranging from 229 kDa to 10 kDa molecular weight. The protein components of p. wsstermani changed chronologically during its developmental period. The 229 kDa band was recognized only in 12-week old worms ($$SEP_{l2}$). 2. Analysis by ELISA showed a significant increase in antibody levels at 3 weeks in infected cats using crude saline extract antigens ($SEP_3,{\;}SEP_5,{\;}SEP_8,{\;}SEP_{l2}$). 3. By EITB using $SEP_3$ and $SEP_5$ infected cats recognisea major protein bands with molecular weight of 60, 35, 28, 25 or 21 kDa at 3~12 weeks of infection, and 3 additional antigens, 19, 13 and 10 kDa, were detected at 8~12 weeks of infections. 4. Using $SEP_8$ 5 antigens, 91, 85, 31, 25 and 21 kDa, were consistently detected by all infected sera tested. In addition, 3 antigens of lg. 13 and 10 kDa were detected at 8~12 weeks of infection. Using $SEP_12$, similar results were obtained with that by using $SEP_8$ and 1 additional antigen of 229 kDa, specifically reacting with the sera from 12 weeks of in(traction, was recognized.

• Journal of Microbiology and Biotechnology
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• v.19 no.8
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• pp.749-759
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• 2009

A survey of Bacillus thuringiensis (Berliner) strains isolated from Spanish citrus orchards has been performed, and the strains were tested for insecticidal activity against the Mediterranean fruit fly Ceratitis capitata (Wiedemann), a key citrus pest in Spain. From a total of 150 environmental samples, 376 isolates were selected, recording a total B. thuringiensis index of 0.52. The collection was characterized by means of phase-contrast microscopy, SDS-PAGE, and PCR analysis with primer pairs detecting toxin genes cry1, cry2, cry3, cry4, cry5, cry7, cry8, cry9, cry10, cry11, cry12, cry14, cry17, cry19, cry21, cry27, cry39, cry44, cyt1, and cyt2. Diverse crystal inclusion morphologies were identified: bipyramidal (45%), round (40%), adhered to the spore (7%), small (5%), and irregular (3%). SDS-PAGE of spore-crystal preparations revealed 39 different electrophoresis patterns. All primer pairs used in PCR tests gave positive amplifications in strains of our collection, except for primers for detection of cry3, cry19, cry39, or cry44 genes. Strains containing cry1, cry2, cry4, and cry27 genes were the most abundant (48.7%, 46%, 11.2%, and 8.2% of the strains, respectively). Ten different genetic profiles were found, although a total of 109 strains did not amplify with the set of primers used. Screening for toxicity against C. capitata adults was performed using both spore-crystal and soluble fractions. Mortality levels were less than 30%. We have developed a large and diverse B. thuringiensis strain collection with huge potential to control several agricultural pests; however, further research is needed to find out Bt strains active against C. capitata.

• The Korean Journal of Zoology
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• v.29 no.2
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• pp.86-106
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• 1986

For the comparative study of protein patterns of the sera of pregnant women, the protein in sera of normal male subjects, non-pregnant women, pregnant women and women delivered of children were analyzed by using the methods of SDS/polyacrylamide gel electrophoresis, two-dimensional electrophoresis and amino acid analysis. The results were as follows: 1. When the protein patterns of sera in normal male ranging from 10, 000 to 110, 000 daltons were compared to non-pregnant women by SDS/polyacrylamide gel electrohporesis, their protein patterns were same each other numerically but bands 3(22, 000 dalton) and 6(39, 000) were less in male than in non-pregnant women quantitatively. When the protein patterns in the pregnant women in which serum were collected two week intervals were compared with non-pregnant women, there was increased or decreased in several bands quantitavely. The protein patterns of sera in pregnant women were compared with those of non-pregnant women; band 3(22, 000) showed similar patterns each other until the 16th week but the quantity of protein was decreased continously from the 18th week to the third trimester of pregnancy. Contary, bands 4(24, 000), 9(69, 000), 10(70, 000), 12(80, 000), 14(86, 000), 15(91, 000) and 16(94, 000) were gradually increased in quantity from the begining of gestation, and band 7(51, 000) was increased until the 32th week of gestation only but somewhat decreased after this time. The quantities of bands 12(80, 000), 15(91, 000) and 16(94, 000) were relatively increased when the protein patterns of delivered women were compared with those of the third trimester of pregnancy. Women who were dilivered female children showed more increase in bands 4(24, 000), 7(51, 000) and 10(70, 000)than one who were delivered male chilren. 2. When the protein patterns of sera in normal males were compared with those of nonpregnant women by two-dimensional electrophoresis, three spots of spot a group were not appeared in the males and the spot c group in the males was less than in the non-pregnant women. In the pregnant women, albumin was significantly decreased during the 10-12 week of gestation but recovered after these times. And spot f(70, 000) was decreased in the 10th week of gestation but increased from this time. 3. Glutamic acid, lysine, aspartic acid, leucine and valine in pregnant women were large in quantity while methionine, isoleucine and glycine were small in quantity by amino acid analysis. The total amino acids were increased remarkably in the second trimester of pregnancy but began to decrease in the third trimester of pregnancy. As mentioned, this present paper deat with that proteins which consist of maternal serum were increased with specific period in pregnancy and that the change of characteristic protein patterns were identified in the serum protein of each trimester in the pregnant women. And furthermore, the study should be preformed for the sex-identification of a fetus in the early pregnancy.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2003.10a
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• pp.95-96
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• 2003

We investigated the proteins that are associated with initiation, maintenance, and termination of diapause in silkworm eggs by means of 2D-gel electrophoresis. 1. Materials : Bombyx mori: unfertilized eggs, diapause eggs, non-diapause eggs, cold-treated egges. 2. Methods: isoelectric focusing, SDS-PAGE, silver staining. (omitted)

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.6
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• pp.885-892
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• 2009

To investigate host defense mechanisms for protecting the mammary gland from mastitis infection, the membrane fraction of mammary tissues from Holstein cows was purified by differential velocity centrifugation, and then the sodium dodecyl sulfate-polyacrylamid gel electrophoresis (SDS-PAGE) separated proteins were identified by ion trap mass spectrometer equipped with a Surveyor high performance liquid chromatography (HPLC) system. A total of 183 proteins were identified. Bioinformatics software was applied to analyse physicochemical characteristics of the identified proteins and to predict biochemical function. These data may provide valuable information to investigate the mechanisms of mammary gland milk secretion and infectious disease, and enable a clear identification of proteins and potential protein targets for therapies.

• BMB Reports
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• v.41 no.3
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• pp.254-258
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• 2008

Lipase was purified from squid (Todarodes pacificus) liver in an attempt to investigate the possibility of applying the enzyme for biotechnological applications. Crude extract of squid liver was initially fractionated by the batch type ion exchange chromatography. The fraction containing lipase activity was further purified with an octyl-Sepharose column. Finally, lipase was purified by eluting active protein from a non-dissociating polyacrylamide gel after zymographic analysis. Molecular weight of the purified enzyme was determined to be 27 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme showed the highest activity at a temperature range of $35-40^{\circ}C$ and at pH 8.0. The activity was almost completely inhibited at 1 mM concentration of $Hg^{2+}$ or $Cu^{2+}$ ion. Partial amino acid sequence of the enzyme was also determined.