• Title/Summary/Keyword: Antigenic difference

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Purification and Characterization of Recombinant Tadpole H-Chain Ferritin in Escherichia coli

  • Chang, So-Ran;Kim, Young-Taek;Kim, Kyung-Suk
    • BMB Reports
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    • v.28 no.3
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    • pp.238-242
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    • 1995
  • The tadpole H-ferritin produced in E. coli was purified and its molecular properties were investigated to obtain information about the contribution of the H-subunit in the reaction of iron core formation. All the expressed subunits were assembled into complete holoprotein in vitro, presumably 24-mer, and the protein was heat-stable. Electron microscopy revealed that the recombinant ferritin forms spherically and contains iron core. No difference was observed in the absorption spectrum of the expressed protein compared to that of the natural ferritin. The Ouchterlony double diffusion of the expressed protein showed that the H-chain ferritin shares an antigenic determinant with natural tadpole ferritin. Rabbit anti-horse spleen ferritin discriminated the H-ferritin from natural ferritin. The rate of ferritin formation by the recombinant H-chain apoferritin was determined to be higher than that shown by natural tadpole ferritin, which consists of H, M and L-subunits. This phenomenon may be caused by the absence of M and L-subunits in the recombinant H-chain apoferritin.

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Investigation of swine toxoplasmosis by Latex agglutination and polymerase chain reaction(PCR) (Latex 응집반응 및 polymerase chain reaction을 이용한 돼지톡소플라즈마병 감염실태 조사)

  • Shim, Hang-Sub;Choi, Gyeong-Muk;Jeon, Oh-Sook;Lee, Su-Jin;Woo, Jong-Tae;Ro, Ki-Woan
    • Korean Journal of Veterinary Service
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    • v.31 no.1
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    • pp.87-91
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    • 2008
  • Between March and October 2007, a total of 516 blood samples from pigs in the Gyeonggi province were examined for seroprevalence against toxoplasmosis by latex agglutination test (LAT) and the detection of antigenic particles among seropositive samples by PCR. In the LAT, 118 (22.8%) were positive, and the unadjusted percentage of seroprevalence rates of breeding and fattening pigs were significant difference. Positive rate (14.1%) in the breeding pigs was much lower than that (27.8%) of the fattening pigs (p<0.001, Pearson's Chi-square test). The antibody detection rate of sows was lower than fattening pigs, i.e., 15.8% (25/158) and 26% (93/358), respectively (P=0.011, Pearson's Chi-square test). Among 118 seropositive samples by LAT, 68 (57.6%) were positive in PCR for the detection of the toxoplasma specific-DNA. There was a statistical difference in the positive PCR reaction between the raising pigs(63/93 67.7%) and sows (5/25, 20%) (P<0.01).

Antibody Responses in Sera of Different Mouse Strains Experimentally Infected with Neodiplostomum seoulense

  • Han, Eun-Taek;Chen, Jun-Hu;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.46 no.4
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    • pp.279-283
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    • 2008
  • To examine humoral immune responses in the host, we measured serum antibody levels in different strains of mice (ICR, BALB/c, and C3H) experimentally infected with Neodiplostomum seoulense. Specific IgG antibody levels were increased remarkably with little difference among 3 strains of mice infected with N. seoulense from day 7 to 35 post-infection. More target proteins of adult parasites reacted with IgG at the time when the worm recovery decreased compared with other times. More than 20 protein bands, from 14 kDa to 94 kDa in size, were separated from the crude antigen of N. seoulense adults by SDS-PAGE, and among them 26, 30, 35, 43, 54, 67, and 94 kDa proteins were the major antigenic proteins. The results suggest that significant IgG antibody responses occur against N. seoulense in mice and this may be related with expulsion of worms.

Development of Bacillus thuringensis var. kurstaki NRD 6-Stu 1 by Site-Directed Mutagenesis (위치지정 변이기법을 이용한 Bacillus thuringiensis var. kurstaki NRD 6-Stu 1의 육성)

  • Lee, Jong-Su
    • The Journal of Natural Sciences
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    • v.6 no.1
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    • pp.41-48
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    • 1993
  • A cloned delta-endotoxin gene from Bacillus thuringiensis var. kurstaki NRD 6 was mutated at N-terminal toxic portion of the protoxin by site-directed mutagenesis. A Stu 1 restriction site was created in protoxin region of the mutant B.t.kurstaki NRD 6-Stu 1 which changed A to C in 177 region of the protoxin(silent mutation). Antigenic property of crystalline inclusion (delta-endotoxin) of the mutant was no difference to intact strain by the western blot analysis and its toxidity threshold value was also showed 0.015~0.030ng against Choristoneura femiferana-1 insect, similar with 0.01~0.024ng of parent strain, but stronger than that of B.t.kurstaki NRD 5 (500~1000ng) and B.t.kurstaki NRD 4(none toxic).

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Immunobiological Studies in Mice Treated with Chemical Carcinogen, 3-Methylcholanthrene : II. Rosette Formation and Natural Killing (NK) Activity of Splenic Lymphocytes (발암제(發癌劑) 3-Methylcholanthrene 투여(投與)마우스에 대(對)한 면역생물학적(免疫生物學的) 연구(硏究) : II. 비장세포(脾臟細胞)의 Rosette형성능(形成能) 및 NK세포(細胞)의 활성(活性))

  • Song, Hee-jong;Kim, Sang-ho;Kim, Jong-myeon
    • Korean Journal of Veterinary Research
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    • v.26 no.1
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    • pp.117-124
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    • 1986
  • The present study was undertaken to evaluate rosette formation and NK activity of splenic lymphocytes in 3-methylcholanthrene (MCA) treated mice. Mice were sensitized iv with 0.1ml of 1% sheep red blood cell (SRBC) suspension were treated with a single ip injection of olive oil alone or with different doses of MCA in oil at various time before or after sensitization, and were challenged at 4 days after SRBC. Rosette formation and NK activity of splenic lymphocytes were measured at 24 hours after challenge. Erythrocyte(E) rosette formation of splenic lymphocytes was significantly depressed in mouse treated with large dose of MCA (5~50mg) regardless of injecting time. But, there was no difference in the response between the treated with small dose of MCA (0.5mg). Whereas erythrocyte-antibody(EA) rosette or erythrocyte-antibody-complement(EAC) rosette forming cells were significantly depressed by MCA. Under small dose of MCA (0.5mg), any difference of NK activity was not observed in all course of injecting time. But, under large dose of MCA, the activity was markedly inhibited to about half the values seen in control and this suppression was transient, resulting that the normal level was reached again 19 days after MCA. These results, which conform with the predictions of immunosuppression hypothesis, suggest that MCA inhibits immunological responses including NK activity and thereby allows the outgrowth of antigenic neoplastic cells.

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Analysis of the Three Dimensional Structure of Envelope Protein of the Japanes encephalitis virus Isolated in Korea (국내에서 분리된 일본뇌염 바이러스의 Envelope Protein의 3차구조 분석)

  • Nam, Jae-Hwan;Chae, Soo-Lim;Kim, Eung-Jung;Yoon, Kyung-Sik;Lee, Ho-Dong;Koh, Hyun-Chul;Cho, Hae-Wol
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.209-216
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    • 1997
  • Three dimensional structures of envelope protein from Korean isolates and Nakayama-NIH strain of Japanese encephalitis virus (JEV) were deduced by a computer program (HyperChem 4.0 Chemplus 1.0) based on the data of the three dimentional structure of Tick-borne encephalitis virus. In the three dimensional structure of envelope protein, neutralizing epitope and T-helper cell recognition site of C-terminal region of Korean isolates were structually similar to those of Nakayama-NIH but the N-terminal region was not. Korean JE isolates were compared with Nakayama-NIH strain by using cross-neutralization antibody test. Neutralizing activities of Korean isolates derived from guinea pigs were higher than those of Nakayama-NIH strain against Korean isolates, although the polyclonal antibody titers of Nakayama-NlH showed 1:160 to 1:640 against Korean isolates. According to the results from three dimentional structures and cross-neutralization analyses, the antigenic difference between Korean JE isolates and Nakayama-NIH strain may be dependent on structural difference of envelope protein.

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A Long-day-stimulus Induced the Expression of c-Fos-like Molecules in the Hypothalamus of Japanese quail

  • Okano, Keiko;Okano, Toshiyuki;Oishi, Tadashi;Fukada, Yoshitaka
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.255-257
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    • 2002
  • In birds, the photoperiodic seasonal breeding involves encephalic photoreception at the initial step of triggering the well-known endocrinal cascade. Especially in Japanese quail (Coturnix coturnixjaponica), the reproductive neuroendocrine function responds to a single long day, and hypothalamic regions are known to be important for the reproductive response. However, little is known about where and how the light and time signals are integrated to detect daylength information and transduced to the endocrinal responses. To gain insights into this issue, we are interested in the c-Fos expression in the hypothalamus of the Japanese quail. Meddle and Follett (1997) previously identified two hypothalamic regions where c-Fos-like immunoreactivities were induced in response to a long day by using an antibody to carboxyl terminal region of human c-Fos (Lys$^{347}$ -Leu$^{367}$ ). In the present study, we used a different anti-c-Fos antibody recognizing a region from Lys$^{128}$ to Ala$^{152}$ of human c-Fos, and found in long-day- stimulated quails many c-Fos-like immunoreactive nuclei localizing within two regions, nucleus anterior medialis hypothalami and nucleus periventricularis hypothalami, which are distinct from those identified in the previous study. Then, we focused on the difference in the cross-reactivities of the antibodies used, and determined the whole coding sequence of quail c-Fos to compare the antigenic sequences of the two antibodies with the amino acid sequence of quail c-Fos. We found that the antibody we used would recognize quail c-Fos more specifically than that used in the previous study.

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Molecular biological characterization of transmissible gastroenteritis viruses isolated in Korea (돼지 전염성 위장염 바이러스(국내분리주)의 분자생물학적 특성 규명)

  • Kwon, Hyuk-moo;Pi, Jae-ho
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.304-313
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    • 1998
  • Sixteen Korean field transmissible gastroenteritis viruses (TGEVs) were isolated using swine testicular cell (STC) and the genomic diversity of them was analyzed. All TGEV isolates produced a typical cytopathic effect in STC and were confirmed as TGEV by immunofluorescence assay using monoclonal antibody against TGEV and PCR using TGEV specific primers. RNAs from TGEV field isolates and vaccine TGEV were extracted and amplified by RT and PCR. The RT-PCR products were digested with selected restriction enzymes and analyzed RFLP patterns. The N-terminal end region of S gene and ORF 3 and 3-1 genes of TGEV amplified by TGEV specific primer pairs seemed to be conserved. Most specific variations were detected in S gene amplified by TGEV 4/6 primer pairs which includes antigenic sites A and D. When the PCR products were treated with Sau3AI and Ssp I, Bvac(vaccine strain), field isolates 133 and 347 were differentiated from Miller and Purdue types. In the case of D5 field isolates, it was classified into Purdue type by Sau 3AI but classified into independent TGEV by Ssp I. Two different TGEV strains from D2 sample were confirmed by plaque purification and RT-PCR-RFLP analysis. To investigate the change occurring in TGEV genome after serial passage, the TGEV P44 strain was passaged through STC. There were specific changes in S gene and a large deletion was observed in ORF 3 and 3-1 genes. These studies showed that a distinct difference in genome exists among TGEV field isolates.

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Diversity of vir Genes in Plasmodium vivax from Endemic Regions in the Republic of Korea: an Initial Evaluation

  • Son, Ui-han;Dinzouna-Boutamba, Sylvatrie-Danne;Lee, Sanghyun;Yun, Hae Soo;Kim, Jung-Yeon;Joo, So-Young;Jeong, Sookwan;Rhee, Man Hee;Hong, Yeonchul;Chung, Dong-Il;Kwak, Dongmi;Goo, Youn-Kyoung
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.149-158
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    • 2017
  • Variant surface antigens (VSAs) encoded by pir families are considered to be the key proteins used by many Plasmodium spp. to escape the host immune system by antigenic variation. This attribute of VSAs is a critical issue in the development of a novel vaccine. In this regard, a population genetic study of vir genes from Plasmodium vivax was performed in the Republic of Korea (ROK). Eighty-five venous blood samples and 4 of the vir genes, namely vir 27, vir 21, vir 12, and vir 4, were selected for study. The number of segregating sites (S), number of haplotypes (H), haplotype diversity (Hd), DNA diversity (${\pi}$ and ${\Theta}_w$), and Tajima's D test value were conducted. Phylogenetic trees of each gene were constructed. The vir 21 (S=143, H=22, Hd=0.827) was the most genetically diverse gene, and the vir 4 (S=6, H=4, Hd=0.556) was the opposite one. Tajima's D values for vir 27 (1.08530, P>0.1), vir 12 (2.89007, P<0.01), and vir 21 (0.40782, P>0.1) were positive, and that of vir 4 (-1.32162, P>0.1) was negative. All phylogenetic trees showed 2 clades with no particular branching according to the geographical differences and cluster. This study is the first survey on the vir genes in ROK, providing information on the genetic level. The sample sequences from vir 4 showed a clear difference to the Sal-1 reference gene sequence, whereas they were very similar to those from Indian isolates.

Evaluation on Immunogenicity and Safety of Avian Influenza Isolate(ADL0401) as a Candidate for the Killed Vaccine against tow-Pathogenic Avian Influenza (약병원성 조류인플루엔자 사독백신개발을 위한 후보주(ADL0401)의 면역 원성 및 안전성 평가)

  • Lee J. S.;Ha D. H.;Kim J. E.;Ha B. D.;Mo I. P.
    • Korean Journal of Poultry Science
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    • v.32 no.2
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    • pp.113-123
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    • 2005
  • Avian influenza (AI) virus (AIV) is distributed worldwide and it has been isolated from various species of wild and domestic birds. AI transfers with high speed and shows diverse pathogenicity syndroms. In Korea, several low Pathogenic AIV, H9N2, have been isolated from the commercial farms with severe decrease of egg production and mortality resulted in severe economic loss since 1996. Therefore, it has been requested to develop AI vaccines to prevent clinical signs and economic losses from the field infection of AIV. To develop a killed vaccine that efficiently prevents low pathogenic AIV (H9N2), evaluation on the pathogenicity and selection of an inactivator for H9N2 is taking place and is being tested safety and immunogenicity of vaccine produced. Based on the pathogenicity test and viral reisolation test, the ADL0401 isolate is the characteristic low pathogenic AIVs and has fairly similar biologic functions compared with MS96 which is the official low pathogenic AIV (H9N2) and one of the predominant AIV isolated from poultry farms in Korea. In antigenicity tests, the ADL0401 and MS96 virus have no significant antigenic difference. In inactivation tests, the ADL0401 isolates can be easily inactivated with $0.1\%$ Formalin at $37^{\circ}C$ within 1 hour with a little decrease of HA titer. The vaccine developed in the present report has no harmful effect on bird and forms good immune capability. Therefore, the isolates, ADL0401 can be used for a killed vaccine which can reduce the clinical signs and viral shedding in the birds infected with H9N2 low pathogenic AIVs.