• 한국응용곤충학회지
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• 제20권2호
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• pp.76-82
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• 1981

아마리리스에 모자익병을 일으키는 병원바이러스의 기주범위, 물리적성질, 혈청반응을 조사하고 바이러스 입자를 전자현미경으로 관찰을 하였다. 바이러스의 지표식물 검정 결과 C. amaranticolor, V. unguiculata, C. sativus G. globosa둥 21종의 CMV감수성식물에 병징이 나타났다. 바이러스의 내열성은 55C(10분간)이며 내희석성은 $10^{-3}$이고 내보존성은 실온에서 2일이었다. 한천내 확산법을 이용한 항혈청반응결과 뚜렷한 반응대가 형성되었으며, 바이러스흡광도는 260nm에서 최고였으며, 245nm에서 최저였다. 전자현미경에 의한 바이러스입자의 검경결과 직경이 90nm내외인 소구형 입자가 관찰되었다. 이상의 결과를 종합하면 아마리리스에 모자익병을 일으키는 것은 CMV에 의한 것으로 보인다.

• 한국응용곤충학회지
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• 제18권1호
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• pp.11-14
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• 1979

짙은 연색의 모자익병징을 나타내는 이병 시금치를 채집하여 Broad bean wilt virus(BBWV)를 분리동정 하였다. 분리된 Broad bean wilt virus (BBWV)를 지표식물에 즙액접종한 결과, 명아주(Chenopodium amaranticolor), 명아주(Chenopodium quinoa) ,잠두(Vicia faba)에서는 접종엽에 국부병반이 나타났고 접종상엽에서는 모자익(전신감염)병징이 나타났으며 꽈리(Physalis floridana), 시금치, 담배 (W.B), 담배 (Nicotiana glutinosa)에는 모자익 병징이 나타났다. 이병시금치로부터 분리한 병원바이러스와 BBWV의 항혈청을 한천내확산법(Agar gel-diffusion test)으로 반응시킨 결과 양성 반응이 나타났다. 이병엽을 Dip법으로 시료를 제작하여 전자현미경에서 검경한 결과 구형의 입자가 관찰되었으며 직경은 25nm이었다. 시금치 재배포장에서 BBWV의 발생분포는 수원, 안양, 대구 진주 등 거의 전국적으로 발생하였다.

• BMB Reports
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• 제31권5호
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• pp.453-458
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• 1998

Human thrombopoietin (hTPO) was expressed to high levels in insect cells using the baculovirus expression system. Full-length hTPO cDNA containing a native signal peptide sequence was amplified by PCR from a human fetal liver cDNA library and cloned into the Autographa californica nuclear polyhedrosis virus (AcNPV) expression vector. Immunoblot analysis with antiserum against hTPO indicated that an approximately 55 kDa protein was produced in recombinant AcNPV infected insect cells. Recombinant hTPO was produced 4-fold higher in Trichoplusia ni (Tn5) cells than in Spodoptera frugiperda (Sf9) cells. with most of the hTPO produced in Tn5 cells secreted into the culture medium. Addition of tunicamycin in the culture medium resulted in the reduction of the size of hTPO to 35-38 kDa, and most of the protein remained within the cell. These results suggest that N-glycosylation of hTPO is required for the secretion of the protein into the culture medium in insect cells. hTPO produced in insect cells induced proliferation and maturation of megakaryocyte progenitors, indicating that it is in a biologically active form.

• BMB Reports
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• 제36권2호
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• pp.214-222
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• 2003

A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$. AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.

• 한국식물병리학회지
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• 제14권3호
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• pp.229-239
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• 1998

Antibodies were raised against fusion proteins of the N-terminus and a region containing the GDNQ (Gly-Asp-Asn-Gln) polymerase motif of the L (polymerase) protein of sonchus yellow net virus (SYNV). Immunoblot analyses using these antibodies revealed the presence of the L protein in purified SYNV preparations and in nuclear extracts from infected tobacco. The serological analyses and detection in a polyacrylamide gels suggested that the L protein is present in at least a 20 fold lower abundance than the G, N, M1 and M2 proteins, and has size corresponding to a molecular weight of over 200 kDa as predicted from nucleotide sequence data. Electron microscopy with gold-labelled antibodies was used to localize the N, M2, and G proteins of SYNV in thin sections of infected tissue. When sections of SYNV-infected tissue were treated with antisera against total SYNV proteins and N protein, gold label could be detected in both the viroplasms and in virus particles. With the anti-M2 protein antiserum, the gold label was strongly localized in the viroplasms but only limited labelling of the virus particle sonly. Limited labelling of the L protein was observed in the viroplasms and the virus particles, presumably because of the low abundance of L protein in the tissues.

• The Plant Pathology Journal
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• 제19권3호
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• pp.166-170
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• 2003

Ninety samples showing mosaic symptoms on soybean (Glycine max) cv. Sukryangputkong were collected from the Cheongsongkun area, Kyungbuk province in Korea. Initially, DAS-ELISA was conducted far detection of Soybean mosaic virus (SMV). Negative samples were chosen at random and mechanically inoculated on soybean cv. Buffalo, which reported not to produce mosaic symptoms when mechanically inoculated with SMV. An isolate of SMV, designated as B-1, from Buffalo showing mosaic and mottle symptoms was used for identification and biological characterization of the causal vim. The purified B-1 isolate had spherical particles of approximately 24nm. It positively reacted with the antiserum against Cowpea mosaic virus (CPMV) but not with Cucumber mosaic virus (CMV) and SMV antisera. CPMV was newly isolated from soybean and had been characterized by host range and by serological and electron microscopic methods. Results of this study suggest that CPMV is the possible cause of mosaic disease in vegetable soybean and that based on sympto-matology, a difference between the typical mosaic and rugose symptoms caused by SMV and CPMV was observed. This is first report of CPMV from soybean in Korea.

• 대한수의학회지
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• 제31권4호
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• pp.397-402
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• 1991

The present experiments were undertaken to produce the standard antiserum for equine blood typing. The following results were obtained through ISO and Hetero Immunizations of the horses whose blood typing was analysed in the Laboratory of Racing Chemistry of Japan. 1. Of the 21 combinations of ISO-immune, 17 horses were produced antibody (about 80%) 2. Antibody titers were increased from early 1 week to late 5 weeks and any antibody titers were not be obtained in spite of the using of adjuvant and 10 repeated injections in the other 4 horses. 3. High antibody titers were obtained within the earliest period in the Dd antigen but were not increased over 32 times in spite of 8~10 repeated injections in the antigen. 4. Antibody were easily produced in the Ca antigen of ISO-Immune but production of antisera were tailed due to abscence of absorbed blood cell. 5. Antibody titers of 1,024 times were obtained through 5 injections in the Ca of HeteroImmune 6. Of the produced 15 antisera (16 system), 13 antigen (5 system) were absorbed.

• 한국수산과학회지
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• 제28권6호
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• pp.753-760
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• 1995

This study was conducted to determine the serum vitellogenin (VTG) concentration changes during the ovulation period in elkhorn sculpin, Alcichthys alcicornis. The results of sepacryl S-300 showed that the molecular weight of VTG could be 380,000. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis may indicate that the purified VTG consists of three subunits with molecular weights of 180,000, 118,000 and 85,000, respectively. Yolk protein purified from the egg extracts was eluted on an equilibrated sephacryl S-300 column, and its molecular weight was estimated 250,000. The precipitation lines of the female serum against the antiserum of the egg extracts were fused completely by immunoelectrophoresis and immunodiffusion analysis. VTG was detected in the serum, and hepatocytes from males injected with $17\beta-estradiol\;(E_2).$ Furthermore, VTG was immunochemically similar to yolk proteins. The concentration of VTG was high before ovulation $(9.80\pm0.81-11.02\pm0.09 mg/ml),$ and then decreased rapidly after ovulation $(less\;than\;6.19\pm0.59 mg/ml).$ This study suggested that VTG was synthesized in the liver by the action of $E_2$ and released to blood, and then incorporated into oocytes.

• International Journal of Oral Biology
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• 제40권4호
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• pp.183-187
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• 2015

The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.

• 한국수산과학회지
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• 제52권6호
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• pp.644-649
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• 2019

Killed vaccines, developed by inactivation with formalin, have been investigated for many fish viruses. In this study, the inactivation of viral hemorrhagic septicemia virus (VHSV) by formalin was investigated based on the infectivity titer. When viral cell culture supernatants were used, the infectivity titer decreased 1,000-fold at 1 d after treatment with 0.1% (v/v) formalin, but was below the detection limit at 7 and 14 d. Moreover, neither the N nor G gene were detectable by RT-PCR immediately after formalin treatment. In western blot analysis, N protein was not detected by rabbit antiserum against VHSV KR-9225 from 2 d after formalin treatment. On the other hand, when we used a virus that was purified and concentrated ~100 times, the infectivity titer was maintained at 10^6.05 TCID₅₀/mL, even at 14 d after formalin treatment, and no change in the viral structural proteins was observed. This study provides important data on the production and use of formalin-inactivated vaccines.