• Title/Summary/Keyword: molecular bands

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Studies on canine babesiosis in Korea I. In vitro isolation and antigenic properties of Babesia gibsoni (개 바베시아병에 관한 연구 I. Babesia gibsoni의 시험관내 분리와 항원성상에 관한 연구)

  • Lee, Ho-kweon;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.681-692
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    • 1996
  • The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

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Effects of Temperature on the Isoenzymes of Peroxidase in Escherichia coli (Escherichia coli의 Peroxidase Isoenzyme에 미치는 온도의 영향)

  • 강사욱;강현삼;홍순우
    • Korean Journal of Microbiology
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    • v.14 no.1
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    • pp.39-47
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    • 1976
  • This experiment was designed to study the effects of temperature on the peroxidase isoenzymes of a mesophilic microorganism, Escherichia coli (grown within biokinetic zone). Optimum temperature for the growth of E. coli was $37{\circ}C.$ Three different temperatures, 20, 30 and $40{\circ}C,$ were selected. And the isoenzyme patterns of peroxidase of E. coli, growth respectively at each temperature, were analysed by disc electrophoresis. The sample of 20.deg.C showed 4 bands, that of 30.deg.C, 5 bands and that of 40.deg.C, 6 bands. Two dark bands (higher molecular weight, 56,000 and 54,000) and two light bands (lower molecular weight, 11,500 and 10,000) were constant at all samples. But two intermediate bands (M.W.44,000 and 34,000) were variable ; at $20{\circ}C,$ no banding pattern, one band 9M.W. 34,000) only at $30{\circ}C,$ and at $40{\circ}C$ two bands were appeared. And the shifts of growth temperatures between 30.deg.C and 40.deg.C showed the alteration of the isoenzyme patterns ; the isoenzyme patterns of the smaple of tmeperature shift from $30{\circ}C$ to $40{\circ}C$ were same as that of 40.deg.C and vice versa.

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A SURVEY OF INTERSTELLAR LINES: RADIAL VELOCITY PROFILES AND EQUIVALENT WIDTHS

  • GALAZUTDINOV GAZINUR
    • Journal of The Korean Astronomical Society
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    • v.38 no.2
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    • pp.215-218
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    • 2005
  • An atlas of high resolution (${\lambda}/{\Delta}{\lambda}$=45,000) profiles of interstellar atomic lines of K I (7665, 7699 ${\AA}$), Na I (D 1, D2), Ca II (H, K), Ca I (4227 ${\AA}$), molecular structures of CH, CH+, CN and the major diffuse interstellar bands at 5780 and 5797 ${\AA}$ based on ${\~}$300 echelle spectra of ${\~}$200 OB stars is presented. Relationships between the reddenings, distances and equivalent widths of NaI, CaII, KI, CH, CH+, CN and diffuse bands are discussed. The equivalent width of K I (7699 ${\AA}$) as well as of CH4300 ${\AA}$ / correlate very tightly with E(B- V) in contrast to the features of neutral sodium, ionized calcium and the molecular ion CH+. The equivalent widths of the Hand K lines of Call grow with distance at a rate ${\~}$250m${\AA}$ per 1 kpc. A similar relation for NaI is much less tight. The strengths of neutral potassium lines, molecular features and diffuse interstellar bands do not correlate practically with distance. These facts suggest that ionized calcium fills the interstellar space quite homogeneously while the other carriers mentioned above, especially K I, CH and these of diffuse bands occupy more and more compact volumes, also filled with dust grains. Apparently the carriers of narrow diffuse bands are spatially correlated with simple molecules and dust grains - all abundant in the so-called 'zeta' type clouds. The same environment seems to be hostile to the carriers of broad diffuse interstellar bands (DIEs) (like 5780 or 6284) and -to a certain extent - also to CaII, NaI and CH+.

Candidate Genes with Ovulation by Differential Display PCR in Small Tail Han Sheep

  • Liu, Shufang;Li, Hongbin;Song, Xuemei;Wang, Aihua;Wei, Caihong;Du, Lixin
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1229-1233
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    • 2006
  • To find the candidate genes concerned with ovulation rate of sheep, Differential Display Reverse Transcription Polymerase Chain Reaction was employed to find the differently expressed cDNA controlling ovulation in the Small Tail Han sheep of polyembryony and in Tan sheep of single birth. Twenty-four primer pairs of three anchored primers and eight arbitrary primers were assembled to amplify the specialized bands from these sheep. Positive cross tests were applied to optimize the ascertainable PCR conditions in which different special bands can be identified by silver strain in one PCR tube. After eliminating the false positive PCR products by Northern hybridization, 24 differential display bands were acquired from the ovary in the Small Tail Han sheep. These EST bands were sequenced and 18 different ESTs were found in which five ESTs had several copies and 13 ESTs had only one copy. Comparing these ESTs with homologous sequences by BLAST in the GenBank, there were six ESTs with known open reading frame (ORF) and function, three ESTs with known ORF and no function, and 9 ESTs without homologous sequence. These ESTs partly represent several genes such as NOS2, tensin, TCRA, CDKN1A, ESR1 and ACTB which express especially in Small Tail Han sheep.

Extraction and Electrophoretic Characterization of Rice Proteins

  • Kim, Mee-sook;Jeong, Yoon-hwa
    • Preventive Nutrition and Food Science
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    • v.7 no.4
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    • pp.437-441
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    • 2002
  • Rice proteins were extracted from brown and milled rice of five varieties: Kwanganbyeo, Daeanbyeo, Daejinbyeo, Surabyeo, Hwaseongbyeo; and their electrophoretic patterns were analyzed by SDS-PAGE. Albumin was extracted with water, globulin with 5% NaCl, prolamin with 70% ethanol, and glutelin with 0.2 M sodium borate buffer (pH 10.0) containing 0.5% SDS, 0.6% $\beta$-mercaptoethanol. The ratios of albumin : globulin : prolamin : glutelin in the brown rice were 10.8~14.1 : 12.4~16.4 : 3.6~5.3 : 68.6~72.8, and in milled rice were 4.4~5.6 : 10.6~12.0 : 3.9~5.4 : 75.7~79.8. In albumin seven major bands were observed with molecular weights ranging from 14.g~96.8 kDa, in globulin four bands with molecular weights in the range of 14.4~56.9 kDa, prolamin had only one band with a molecular weight of 14.4 kDa, and glutelin had four bands with molecular weights of 14.4 ~ 57.4 kDa. There were no differences in electrophoretic patterns between rice varieties or between brown and milled rice.

Molecular Weight Estimation and Protein Patterns of Panax ginseng, Codonopsis lanceolata and Platycodon grandiflorum by Polyacrylamide gel Electrophoresis (인삼, 사삼 및 도라지의 전기영동에 의한 단백질 패턴과 분자량)

  • Ahn, Sang-Deuk;Chung, Chan-Moon;Kwon, Woo-Saeng;Son, Eung-Ryong;Lee, Yong-Sae;Yoon, Kyeung-Eun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.4
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    • pp.352-358
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    • 1985
  • This study was carried out to identify the protein patterns and to estimate molecular weights of the ginseng and other plants. Polyacrylamide gradient 2-30% gels for identification of protein patterns and 11% SDS polyacrylamide gels for molecular weight estimation of various plants were used. Proteins extracted with distilled water and phosphate buffer were most clearly separated. Although minor bands were a little different according to kinds of buffer, major bands showed similar pattern. Major protein bands were two bands of 45,000, 66,000 in Panax ginseng, 3 bands of 32,000-39,000 in Codonopsis lanceolata and one double band around 39,000 kd in Platycodon grandiflorum.

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Molecular Dyamics Simulation and Far Infrared Measurements of $Ba_{0.6}K_{0.4}BiO_3$

  • Lee, C.Y.;Song, Ki.Y.;Sperline, R.P.
    • Korean Journal of Materials Research
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    • v.6 no.6
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    • pp.555-560
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    • 1996
  • The vibrational behavior and the molecular dynamics of the high Tc superconductor Ba0.6K0.4BiO3 have been studied experimentally and by atomistic computer simulation methods. For Ba0.6K0.4BiO3, the vibrational spectrum is dominated by oxygen ion modes from 150cm-1 to 820cm-1 including infrared absorption bands at 330, 480, 640 and 830cm-1including infrared absorption bands at 330, 480, 640 and 830cm-1at room temperature. Band assignments are discussed in relation to those bands predicted by simulations, and the infrared and Raman measurements reported in the literature.

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Relationship between Infrared Peak Maximum Position and Molecular Interactions

  • Ryu, Soo-Ryeon;Noda, Isao;Jung, Young-Mee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.11
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    • pp.4011-4015
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    • 2011
  • We explored the interpretation of the well-accepted correlation between the apparent peak maximum position shift and extent of molecular interactions, like hydrogen bonding and dipole-dipole interactions, based on the overlapped multiple band model. The simulation of two overlapped Lorentzian bands was carried out to interpret how the maximum position of a composite peak relates to the relative contributions of two species representing the different levels of molecular interactions, i.e., free (or very weekly bound) vs. strongly bound. To demonstrate the validity of our interpretation of the origin of the peak position shift, the temperaturedependent IR spectra of ethylene glycol were also analyzed. It was found through the analysis of simulated and experimental spectra that the apparent peak shift in certain case can be safely interpreted as the measure of the strength of hydrogen bonding. The result of this study gives a new insight to interpret molecular interactions probed by vibrational spectroscopy.

DNA Microarray Probe Preparation by Gel Isolation Nested PCR

  • Wang, Hong-Min;Ma, Wen-li;Huang, Hai;Xiao, Wei-Wei;Wang, Yan;Zheng, Wen-Ling
    • BMB Reports
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    • v.37 no.3
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    • pp.356-361
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    • 2004
  • To develop a simplified method that can rapidly prepare DNA microarray probes in a massive scale, a lambda phage genomic DNA-fragments library was constructed for the microarray-probes collection. Four methods of DNA band recovery from the first PCR products were tested and compared. The DNA microarray probes were collected by a novel method of nested PCR that was mediated by gel isolation of the first PCR products. This method was named GIN-PCR. The probes that were prepared by this GIN-PCR technique were used as subjects to fabricate a DNA microarray. The results showed that a wooden toothpick was superior to the other 3 methods, since this technique can steadily transfer the DNA bands as the template of the second PCR after the first PCR. A group of probes were successfully collected and DNA microarrays were constructed using these probes. Hybridization results demonstrated that this technique of DNA recovery and probe preparation was rapid, efficient, and effective. We developed a cost-effective and less labor-intensive method for DNA microarray probe preparation by nested PCR that is mediated by wooden toothpick transfer of the DNA bands in the gel after electrophoresis.

Purification and Characterization of Phytoferritin

  • Oh, Suk-Heung;Cho, Sung-Woo;Kwon, Tae-Ho;Yang, Moon-Sik
    • BMB Reports
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    • v.29 no.6
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    • pp.540-544
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    • 1996
  • Ferritins from germinated pumpkin seeds were isolated by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, and gel filtration chromatographies on Sephacryl S-300 and Sephadex G-100. Pumpkin ferritin contains less iron than soybean ferritin. Pumpkin ferritin cross-reacted with anti-soybean ferritin antiserum made in rabbit, and showed two distinct antibody reactive bands, both of equal intensity. The pumpkin ferritins corresponding to the two bands were separable by centrifugation in a sucrose gradient (20~50%). The molecular weights of the native pumpkin ferritins based on the estimation of sucrose gradient centrifugation, gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be: 530~580 KD (the large molecular weight pumpkin ferritin) and 330-360 KD (the small molecular weight pumpkin ferritin) The large molecular weight pumpkin ferritin contains less iron. Both pumpkin ferritins cross-reacted with anti-soybean ferritin antibody with a spur formation suggesting partial antigenic recognition.

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