• BMB Reports
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• 제34권1호
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• pp.33-38
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• 2001

The human PTK6 (also known as Brk) polypeptide, which is deduced from its full-length cDNA, represents a non-receptor protein tyrosine kinase (PTK). It contains SH3, SH2, and tyrosine kinase catalytic domains that are closely related to Src family members. We generated an antihuman PTK6 antibody by immunizing rabbits with a PTK6-specific oligopeptide conjugated to BSA, which corresponds to 11 amino acid residues near the C-terminus. An immunoblot analysis with the antibody detected an expected 52-kDa band in various mammalian transformed cell lines. Immunoprecipitation and immunoblot analyses demonstrated that PTK6 is phosphorylated on the tyrosine residues) and interacts with approximately a 23-kDa tyrosine-phosphorylated polypeptide (most likely a substrate of PTK6) in breast carcinoma T-47D cells. An immunofluorescence analysis demonstrated that PTK6 is localized throughout the cytoplasm of T-47D cells. These results support a possible role for PTK6 in the intracellular signal transduction through tyrosine phosphorylation.

• 대한임상검사과학회지
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• 제39권3호
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• pp.201-209
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• 2007

Among the inhalant and foods which caused allergy, which is increasing nowaday, this research has a purpose of studying allergy-causing materials (allergen) which most frequently cause allergy. In order to conduct this research, we selected 100 hundred patients (M : F = 65 : 35) from in Kwangju Christian Hospital, who were tested through allergy multiple antigen simultaneous test (MAST) immunoblot assay (RIDA$^{(R)}$ X-Screen; r-Biopharm Co., Darmstadt, Germany). According to the test, the total IgE positive rate (above Class 2) from the inhalant is 58.3%, and that from food is 35%, respectively. This research showed that the first most allergen was the tick (Derm. farinae/Derm. pteronyssinus), the second most was housedust, and the third was the hair of pets such as cats and dogs; 65%, 35%, 26.3% in inhalant panel, and 52.5%, 40%, 50% in food panel, respectively. Thease allergens newly added to inhalant and food panels were not contributable to the detection of specifical allergen. We needs further improvement to be used as primary screening or quantitative test for allergy.

• 미생물학회지
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• 제32권1호
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• pp.34-39
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• 1994

Schwanniomyces castellii의 제놈 DNA로 제조된 유전자 은행으로부터 cloning된 ${\alpha}$-amylase 유전자가 Sacchromyces cerevisiae에서 발현되었다. Cloning된 삽입 DNA 절편의 크기는 약 5.0 kb이었고, Southern 및 immunoblot 분석 결과 cloning된 ${\alpha}$-amylase 유전자가 Sch. Castellii로부터 유래되었음이 확인되었다. S. cerevisiae SHY3 형질전환체에서 Sch. Castellii ${\alpha}$-amylase 유전자발현은 모균주에 비해 낮았으나, 단백질의 분자량 및 효소의 성질은 Sch. Castellii에서 분리한 ${\alpha}$-amylase의 그것과 차이가 없었다.

• IMMUNE NETWORK
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• 제6권3호
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• pp.123-127
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• 2006

Background: Caveolin-1 is a principal component of caveolae membranes in vivo. Although expression of caveolae structure and expression of caveolin family, caveolin-1, -2 and -3, was known in chondrocytes, the functional role of caveolae and caveolins in chondrocytes remains unknown. In this study, we investigated the role of caveolin-1 in articular chondrocytes. Methods: Rabbit articular chondrocytes were prepared from cartilage slices of 2-week-old New Zealand white rabbits by enzymatic digestion. Caveolin-1 cDNA was transfected to articular chondrocytes using LipofectaminePLUS. The cyclooxygenase-2 (COX-2) expression levels were determined by immunoblot analysis, immunostaining, immunohistochemistry, and prostaglandin $E_2\;(PGE_2)$ assay was used to measure the COX-2 activity. Results: Ectopic expression of caveolin-1 induced COX-2 expression and activity, as indicated by immunoblot analysis and $PGE_2$ assay. And also, overexpression of caveolin-1 stimulated activation of p38 kinase and ERK-1/-2. Inhibition of p38 kinase and ERK-1/-2 with SB203580 and PD98059, respectively, led to a dose-dependent decrease COX-2 expression and $PGE_2$ production in caveolin-1-transfected cells. Conclusion: Taken together, our data suggest that ectopic expression of caveolin-1 contributes to the expression and activity of COX-2 in articular chondrocytes through MAP kinase pathway.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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• pp.446-446
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• 2000

Specific polyclonal and/or monoclonal antibodies (MAbs) to immunoglobulins (Igs) and their subunits have proved to be valuable tools in immunological research and in immunological assays. In this study, we developed and characterized MAbs against flounder serum Igs. To obtain the pure flounder serum Igs, mouse IgG (mIgG) was immunized to flounder. Flounder Igs were purified by using mIgG-agarose affinity column chromatography. The structure of purified flounder Ig was observed, on denatured SDS-PAGE, to be composed of two heavy chains (77 and 72 kd) and two light chains (28 and 26 kd). MAbs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously primed with the flounder Igs. Finally, three hybridoma clones, FIM 511, FIM 519 and FIM 562 were established to recognize both 2 heavy chains, 26 kd of light chain and 28 kd of light chain, respectively. On the other hand, the flounder immune sera collected on the weekly basis were tested on ELISA and immunoblot analysis whether boosting effect is present in flounder humoral immune system. As a result, the secondary immune response in flounder was ascertained on ELISA, but not on immunoblot analysis. Further, we observed an alteration of serum protein levels following immunization. Our MAbs and basic information on flounder humoral immune system obtained in this study will be helpful to control and monitor the efficiency of fish vaccines and therapeutic process of flounder diseases.

• Parasites, Hosts and Diseases
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• 제38권3호
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• pp.145-150
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• 2000

Antigenic components in the crude extracts of Spirometra mansoni plerocercoid were analyzed in early experimental infections and in IgG subclass observed in clinical sparganosis. By IgG immunoblot, sera obtained serially from experimental mice, fed 5 spargana each, were reacted with the crude extracts. Protein bands at 36-26 kDa and 103 kDa showed positive reactions since two weeks after infection. In a differential immunoblot, in which a monospecific antibody against sparganum chymase at 36 kDa was pre-treated, the reactions at 36-26 kDa disappeared, indicating that the sparganum chymase and its degradation products invoked IgG antibody reactions. When 69 patients sera of human sparganosis were examined for their IgG subclass responses, IgG4 levels showed the highest reaction which was followed by IgG 1 The IgG4 antibody also reacted mainly with 36-31 kDa protease. These results indicate that 36 kDa chymase of 5. nansoni plerocercoid is the main antigenic component inducing Ige antibody response in early stage of experimental sparganosis and for specific IgG subclass reactions in human sparganosis.

• 식물조직배양학회지
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• 제27권3호
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• pp.239-243
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• 2000

위염과 위궤양을 일으키는 Helicobacter pylori는 그 감염 환자에서 강한 항체형성을 유도하는 urease를 생산한다. 식물체를 이용하여 H. pylori에 대한 식용백신 모델을 제조하기 위하여 H. pylori의 urease 유전자를 지니고 있는 pH808 plasmid로부터 750bp의 ureA DNA를 PCR에 의해서 증폭한 후 이를 담배식물에서 발현이 되도록 조작하였다. 재분화된 형질전환 식물체로부터 ureA 유전자의 도입과 mRNA발현 및 단백질합성을 분석하였다. CaMV 35S promoter에 의한 ureA 유전자의 발현은 SDS polyacrylamide 전기영동 및 immunoblot실험에서 박테리아로부터의 재조합 단백질과 같은 크기의 30 kDA UreA 단백질이 생산되었음을 확인할 수 있었다.

• Food Science and Biotechnology
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• 제15권4호
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• pp.612-616
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• 2006

The protective effect of water extract of ash tree leaves (ALE) against oxidative damages was investigated in paracetamol-induced BALB/c mice. Biochemical analysis of anti-oxidative enzymes, immunoblot analyses of hepatic cytochrome P450 2El (CYP2E1), and the gene expression of tumor necrosis factor (TNF-${\alpha}$) were examined to determine the extract's protective effect and its possible mechanisms. BALB/c mice were divided into three groups: normal, paracetamol-administered, and ALE-pretreated groups. A single dose of paracetamol led to a marked increase in lipid peroxidation as measured by malondialdehyde (MDA). This was associated with a significant reduction in the hepatic antioxidant system, e.g., glutathione (GSH). Paracetamol administration also significantly elevated the expression of CYP2E1, according to immunoblot analysis, and of TNF-${\alpha}$ mRNA in liver. However, ALE pretreatment prior to the administration of paracetamol significantly decreased hepatic MDA levels. ALE restored hepatic glutathione and catalase levels and suppressed the expression of CYP2E1 and TNF-${\alpha}$ observed in inflammatory tissues. Moreover, ALE restored mitochondrial ATP content depleted by the drug administration. These results show that the extract of ash tree leaves protects against paracetamol-induced oxidative damages by blocking oxidative stress and CYP2E1-mediated paracetamol bioactivation.

• Parasites, Hosts and Diseases
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• 제34권4호
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• pp.247-254
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• 1996

임상증상 없이 검변을 통하여 검출된 이질아메바 포낭양성자를 어떻게 다루어야 할지 오랫동안 논란이 있었으나 이중에서 병원성이 강한 이질아메바(E. histolytica)만 치료하고 병원성이 없는 동형아메바(E. dispar)는 치료하지 않는 것으로 의견이 모아져 있다. 따라서 이 두 종을 감별하는 것이 실제적으로 중요하게 되었고 이를 위하여 동위효소 분석 유전자 분석. 혈청학적인 검사가 가능하다. 그러나 앞의 두 분석법은 원충을 배양하여야 가능하기 때문에 적용하기 어려운 경우가 많아 혈청학적인 감별이 실제적으로 유용하다. 이 연구는 우리 나라에서 무균배양에 성공한 이질 아메바 YS-27의 항원성을 확인된 아메바 감염자 혈청과 면역이적법으로 검증하고. 이를 이용하여 무증상 포낭배출자에서 두 종의 감염을 감별하고자 하였다. 시스테인계 단백질분해효소 억제제인 E64를 첨가한 이질아메바의 항원은 117. 110. 99. 68. 66. 60. 54. 52 46. 45 kDa 분획이 이질아메바 감염이 확인된 사람의 혈청내 IgG 항체와 반응하였고 동형아메바 대장아메바 Beastnystis 및 대조군의 혈청과는 117 kDa 분획이 미약하게 반응하였다. 포낭양성자의 혈청내 IgA 항체와 많은 분획이 반응하였으나 두 종 양성자간에 차이가 없었고 혈청내 IgM과 IgE 항체와 반응하는 분획이 없었다. 동형아메바의 항원은 두 종 양성자 혈청내 모든 군의 항체와의 반응에서 두종을 감별하지 못하였다. 따라서 이질아메바의 항원과 혈청내 IgG 항체 반응이 두 아메바 감염 을 감별할 수 있었다. 증상이 없는 포낭배출자 24명의 혈청을 면역이적 반응시킨 결과 2명에서 46 과 45 kDa 분획을 제외한 다른 모든 분획과 반응하여 이질아메바 감염자로 확인하였고 22명은 동 형아메바 양성자로 확인하였다. 이 자료를 토대로 하면 우리 냐라에서 검출되는 단순 포낭배출자 의 90% 이상은 이질아메바가 아니라 동형아메바 감염자이고. 이에 따라서 포낭양성자 중 환자가 많지 않은 사실을 설명할 수 있을 것이다. 이질아베바 조항원을 이용한 면역이적법은 증상이 없는 아메바 포낭배출자 중 치료를 꼭 필요로 하는 사람을 찾아내는 방법으로 유용함을 확인하였다.

• Parasites, Hosts and Diseases
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• 제32권1호
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• pp.35-42
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• 1994

간흡충 특이 IgG 항체와 간흡충 분비배설항원간의 면역반응이 간흡충 감염과 치료에 의해 어떻게 변화되고 치료 후 소실되는 것이 있는지 관찰하였다. 간흡충 피낭유충 150, 450개씩 토끼에 감염시키고 감염대조군은 22개월. 프라지콴텔 치료군은 감염 4개월에 투약하였으나 완치되지 않아 이후 $4\frac{1}{2}$개월에 재투약하여 완치시킨 후 13개월까지 관찰하였다. 간흡충 분비배설항원은 SDS-PAGE에서 114, 102, 94, 80, 66, 63, 61, 59, 54, 52, 50, 47, 45, 42, 40, 38, 34, 33, 30, 27, 25, 23, 20, 19, 18, 17, 16, 12.5, 12, 11.5 kDa 단백질 구성이었다. 이중 항 원 단백질은 감염혈청과 치료후 혈청으로 Immunoblot할 경우 66, 63(A군) 54-38(B군) 34. 33(C군), 30-23(D군) 20-14(I군) 12, 12.5, 11.5(K군) kDa 단백질이었다. 특히 33. 27, 13. 12.5 kDa 항원은 감염기간 동안 가장 강한 면역반응을 나타낸 항원이었고 피낭유층 감염강도에 의한 차이는 없었다. 또한 투약은 했으나 완치되지 않은 4개월여 동안 면역반응의 변화는 없었다. 치료후 면역반응은 33 27 kDa 항원의 경우 환치후 13개월까지 지속되었고 치료전과 정도 차이가 없었다. 그러나 13, 12.5 kDa 항원의 면역반응은 완치후 1개월부터 약화되기 시작하여 6 개훨이면 소멸되는 양상이 뚜렷하였다. 12.5 MDa 항원은 SDS-PAGE상 단백질 함량이 많고 간흡충 감염의 강한 항원이면서 치료 후에는 특이항체와의 면역반응이 치료전에 비해 현격히 감소되므로 간흡충 현증 감염만을 혈청학적으로 진단해 줄 수 있는 항원으로 판단하고 간흡충 'K2-항원'이라 명명하였다.