• Title/Summary/Keyword: 2-D gel electrophoresis

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Electrophoretic Study of Semisulcospira gottschei in Korea (한국산(韓國産) Semisulcospira gottschei의 전기영동적(電氣泳動的) 연구(硏究))

  • Kim, Chang Han
    • The Korean Journal of Malacology
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    • v.2 no.1
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    • pp.30-34
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    • 1986
  • Genetic variations of Semisulcospjra gottschei in Korea were investigated by means of starch gel-electrophoresis. The results are as follows; 1) Eight loci of ${\alpha}$-Gpd, Mdh-2, Pept-1, Pgm-2, Gp-1, Gp-2, Gp-3 and Gp-4 were monomorphic, and 8 loci of Mpi, Mdh-1, Pgi, Got, Pept-2, Pept-3, Pgm-1 and Sdh showed genetic variations. 2) Means of allele per locus ($\bar{A}$) and polymorphism (P) were revealed 0.64 and 0.44, respectively. The value of heterozygosity ($H_G$) showed a little more than heterozygosity $H_D$. ($H_D$=0.09, $H_G$=0.13) 3) Genetic variation appears to be due to loci of Got, Pept, Pgi, and Sdh, all of which of the $H_D$ values are high.

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Electrophoretic Behaviors of α-Lactalbumin and β-Lactoglobulin Mixtures Caused by Heat Treatment

  • Lee, You-Ra;Hong, Youn-Ho
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1041-1045
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    • 2003
  • In order to study the reaction behaviors of bovine $\alpha$-lactalbumin ($\alpha$-La), $\beta$-lactoglobulin ($\beta$-Lg), and their mixtures during heat treatment, samples were analyzed using native-polyacrylamide gel electrophoresis (Native-PAGE), sodium dodecylsulfate (SDS)-PAGE, and two-dimensional (2-D)-PAGE. The electrophoresis demonstrated that the loss of native-$\alpha$-La increased as temperature increased, and that the loss of apo-$\alpha$-La was slightly higher than that of holo-$\alpha$-La. The tests also showed that during heat treatment, a mixture of $\alpha$-La and $\beta$-Lg was less stable than $\alpha$-La alone. As such, it was assumed that $\beta$-Lg induced holo-$\alpha$-La to be less stable than apo-$\alpha$-La during heat treatment. The reaction behavior of $\alpha$-La (holo-, apo-form) during heat treatment showed similar patterns in the 2-D-PAGE electropherogram, but the mixture of $\alpha$-La and $\beta$-Lg created new bands. In particular, the results showed a greater loss of native $\alpha$-La in the holo-$\alpha$-La and $\beta$-Lg mixture than in the apo-$\alpha$-La and $\beta$-Lg mixture. Thus, it can be concluded that the holo-$\alpha$-La and $\beta$-Lg mixture was more intensively affected by heat treatment than other samples, and that free sulphydryl groups took part in the heat-induced denaturation.

Studies on hematologic values and types of blood protein in Jindo dogs II. Types of blood protein of Jindo dogs (진도견(珍島犬)의 혈액상(血液像)과 혈액단백질형(血液蛋白質型)에 관(關)한 연구(硏究) II. 진도견(珍島犬)의 혈액단백질형(血液蛋白質型))

  • Kim, Woo-kwon;Han, Bang-keun;Kim, Ja-suk
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.299-305
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    • 1988
  • The phenotypes of hemoglobin, albumin and transferrin of 3U2 Jindo dogs in Jindo area were studied by starch gel electrophoresis for hemoglobin and albumin, and by polyacrylamide gradient gel electrophoresis for transferrin. The results obtained were as follows: 1. In the hemoglobin phenotypes, three phenotypes, HbAA, HbAB and HbBB, which were controlled by two allelic genes, $Hb^A$ and $Hb^B$, were observed and their frequencies of appearance were 1.65%, 10.60% and 87.75% respectively. The distribution of gene frequency was calculated as 0.0695 in $Hb^A$ and 0.9305 in $Hb^B$. 2. In the albumin phenotypes, three phenotypes, Alb FF, Alb FS and Alb SS, which were controlled by two allelic genes, $AIb^F$ and $AIb^S$ were observed and their frequencies of appearance were 12.59%, 25.56% and 61.85% respectively. The distribution of gene frequency was calculated as 0.2537 in $AIb^F$ and 0.7463 in $AIb^S$. 3. Analysis of transferrin phenotypes showed 6 different types which were controlled by three allelic genes, $Tf^B$, $Tf^C$ and $Tf^D$ and their frequencies of appearance were 54.04% in TfBB, 17.54% in TfBC, 9.82% in TfBD, 8.07% in TfCC, 7.37% in TfCD and 3.16% in TfDD. The distribution of gene frequency was calculated as 0.6772 in $Tf^B$, 0.2053 in $Tf^C$ and 0.1175 in $Tf^D$.

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Expression of Acid Stress-Induced Proteins of Streptococcus mutans Isolated from Korean Children with Caries (한국인 우식아동으로부터 분리한 Streptococcus mutans의 내산성 단백질의 발현)

  • Kang, Kyung-Hee;Nam, Jin-Sik;Jin, Ing-Nyol
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.7
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    • pp.1766-1772
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    • 2009
  • In this study, we are interested in comparing the protein profiles of acid-shocked and control cells of S. mutans isolated from Korean children with caries. The results of 2D gel electrophoresis showed that twelve proteins are up-regulated when the cells were grown under 20 mM lactic acid stress in the exponential phase. Up-proteins under acid stress were estimated a major key of the survival and proliferation of S. mutans in low pH environments. These proteins are estimated generally associated with three biochemical pathways: glycolysis, alternative acid production and branched-chain amino acid biosynthesis.

Identification of a Marker Protein for Cardiac Ischemia and Reperfusion Injury by Two-Dimensional Gel Electrophoresis and Matrix-Assisted Laser Desorption Ionization Mass Spectrometry

  • Lee, Young-Suk;Kim, Na-Ri;Kim, Hyun-Ju;Joo, Hyun;Kim, Young-Nam;Jeong, Dae-Hoon;Cuong, Dang Van;Kim, Eui-Yong;Hur, Dae-Young;Park, Young-Shik;Hong, Yong-Geun;Lee, Sang-Kyung;Chung, Joon-Yong;Seog, Dae-Hyun;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.4
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    • pp.207-211
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    • 2004
  • The purpose of the present study was to evaluate the expression of cardiac marker protein in rabbit cardiac tissue that was exposed to ischemic preconditioning (IPC), or ischemiareperfusion injury (IR) using two-dimensional gel electrophoresis (2DE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). We compared 2DE gels of control (uninjured) cardiac tissue with those of IPC and IR cardiac tissue. Expression of one protein was detected in IR heart tissue, however the protein was not detected in the samples of control and IPC tissue. To further characterize the detected protein molecule, the protein in the 2D gel was isolated and subjected to trypsin digestion, followed by MALDI-MS. The protein was identified as myoglobin, which was confirmed also by Western blot analysis. These results are consistent with previous studies of cardiac markers in ischemic hearts, indicating myoglobin as a suitable marker of myocardial injury. In addition, the present use of multiple techniques indicates that proteomic analysis is an appropriate means to identify cardiac markers in studies of IPC and IR.

Exploitation of the Dose/Time-Response Relationship for a New Measure of DNA Repari in the Single-Cell Gel Electrophoresis (Comet) Assay

  • Kim, Byung-Soo;Edler, Lutz;Park, Jin-Joo;Fournier, Dietrich Von;Haase, Wulf;Sautter-Bihl, Mare-Luise;Hagmuller, Egbert;Gotzes, Florian;Thielmann, Heinz Walter
    • Toxicological Research
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    • v.20 no.2
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    • pp.89-100
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    • 2004
  • The comet assay (also called the single-cell gel electrophoresis assay) has been widely used for detecting DNA damage and repair in individual cells. Since the conventional methods of evaluating comet assay data using frequency statistics are unsatisfactory we developed a new quantitative measure of DNA damage/repair that is based on all information residing in the dose/time-response curves of a comet experiment. Blood samples were taken from 25 breast cancer patients before undergoing radiotherapy. The comet assay was performed under alkaline conditions using isolated lymphocytes. Tail DNA, tail length, tail moment and tail inertia of the comet were measured for each patient at four doses of $\gamma$-rays (0, 2, 4 and 8 Gy) and at four time points after irradiation (0, 10, 20 and 30 min) using 100 cells each. The resulting three-dimensional dose-time response surface was modeled by multiple regression, and the second derivative, termed 2D, on dose and time was determined. A software module was programmed in SAS/AF to compute 2D values. We applied the new method successfully to data obtained from cancer patients to be assessed for their radiation sensitivity. We computed the 2D values for the four damage measures, i.e., tail moment, tail length, tail DNA and tail inertia, and examined the pairwise correlation coefficients of 2D both on the log scale and the unlogged scale. 2D values based on tail moment and tail DNA showed a high correlation and, therefore, these two damage measures can be used interchangeably as far as DNA repair is concerned. 2D values based on tail inertia have a correlation profile different from the other 2D values which may reflect different facets of DNA damage/repair. Using the dose-time response surface, other statistical models, e.g., the proportional hazards model, become applicable for data analysis. The 2D approach can be applied to all DNA repair measures, Le., tail moment, tail length, tail DNA and tail inertia, and appears to be superior to conventional evaluation methods as it integrates all data of the dose/time-response curves of a comet assay.

Purification and Characterization of A Thermotolerable Restriction Endonuclease from Streptomyces violochromogenes D2-5

  • Yun, Mi-Sub;Hwang, Hye-Yeon;Bae, Moo
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.269-273
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    • 1995
  • A thermotolerable restriction endonuclease. Svil, found in Streptomyces violochromogenes D2-5 was purified. For the purification, streptomycin sulfate and ammonium sulfate precipitation was used. Ph osphocellulose P-ll, DEAE-Cellulose and Sephacryl-S200 HR colum chromatography were also performed. The purified enzyme was found to be homogeneous and the molecular weight of the enzyme estimated by polyacrylamide gel electrophoresis containing 0.1$%$ SDS was about 32, 000 daltons. The recognition sequence and cleavage site of the enzyme were determined to be $5^1$-$TT\downarrow CGAA$-$3^1$ which is the same sequence as that of Asull. Unlike Asull, however, the Svil shows high thermal stability.

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HABIT : Cancer Diagnosis System (HABIT : 질병 진단 시스템)

  • Kim, Gi-Seong;On, Seung-Yeop;Gang, Gyeong-Nam
    • Proceedings of the KIEE Conference
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    • 2003.11c
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    • pp.898-902
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    • 2003
  • In this paper we proposes a new technique for identification of breast cancer by classification of proteome pattern generated from 2-D polyacrylamide gel electrophoresis (2-D PAGE) and development of cancer diagnosis system : HABIT. Proteome patterns reflect the underlying pathological state of a human organ and it is believed that the anomalies or diseases of human organs are identified by the analysis or classification of the patterns. Proteome patterns consist of quantitative information of the spots such as their size, position, and density in the proteome image produced from 2-D PAGE, for the Image mining of proteome pattern, SVM(support vector machine) and GA(genetic algorithm) are used to generate a decision model for the identification of breast cancer The decision model was then used to classify an independent set of test proteome patterns into the affecter and unaffecter classes. The proposed technique was tested by actual clinical test samples and showed a good performance of a hit ratio of 90%.

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SAFETY EVALUATION OF ADENOVIRUS-MEDIATED P16 GENE TRANSFER BY USING MICROARRAY AND 2D/MALDI-TOF

  • Park, Misun;Hoil Kang;Jaehee Pyo;Sinae Lim;Seungwan Jee;Miok Eom;Taikyung Ryeom;Kim, Okhee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.196-196
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    • 2002
  • p16INK4a tumor suppressor gene transfer in the non-small cell lung cancer cells by transduction of recombinant adenovirus (Ad5CMV-p16) resulted in significant inhibition of cancer cell growth (Anticancer Res., 1998, 18:3257-3261). As a safety concern, we have investigated gene and protein expression after transduction of adenoviral vector (Ad5CMV-p16) in human non-small cell lung cancer (A549) cells by using microarray and 2D gel electrophoresis/ MALDI-TOF.(omitted)

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Analysis of Diapause-Associated Changes in Silkworm Egg Proteins

  • Go, Hyun-Jeong;Hwang, Jae-Sam;Kim, Young-Tae;Kim, Hyun-Su;Kang, Seok-Woo;Chang, Jong-Su;Lee, Sang-Mong;Lee, Bong-Hee;Seong, Su-Il
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.1
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    • pp.117-121
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    • 2004
  • The patterns of diapause-associated proteins of silkworm eggs were analyzed by two-dimensional (2-D) gel electrophoresis. Among the hundreds of spots on the 2-D gels, at least two proteins were considered to be associated with diapause. A protein, spot 4, with an approximate molecular weight of 38 kDa and pI 6.1 was observed in the HCI-treated, cold-treated, and diapause eggs, respectively. Spot 4 was undetectable in unfertilized eggs and non-diapause eggs at two days after oviposition, suggesting that this protein may be associated with the entrance to diapause. A protein, spot 11, with an approximate molecular weight of 21 kDa and pI of 61 was detected in the unfertilized, HCl-treated, and cold-treated eggs, respectively, after oviposition by normal moths. In diapausing eggs, a protein corresponding to spot 11 was observed in 3-, 5-, and 30-day-old eggs, while the protein was not detected one-day-old eggs. The protein corresponding to spot 11 was not detected in unfertilized and non-diapause eggs obtained from subesophabeal ganglion (SG)-extirpated moths either. Spot 11 was also considered to be a diapause specific protein, which occurred at only early embryonic stage under the control of diapause-downregulated gene.