피르빅산 탈수소 효소(돼지 신장)의 순수 정제

권무식,홍성렬 成均館大學校 科學技術硏究所 1990 論文集 Vol.41 No.1

Pyruvate dehydrogenase(El) composed of two nonidentical subunits a & b has been isolated from Pyruvate dehydrogenase complex(PDC) (porcine kidney). Essential steps in this purification process are involved in fractionation of PDC component by sodium dodecyl sulfate -polyacrylamide gel electrophoresis (SDS -PAGE), electroelution of E1 (a, b) using dialysis membrane, and ethanol precipitation of eluate. Anti-El antiserum has been raised in a rabbit against El(a, b) purified from porcine kidney. The anti-El antiserum is immunoreacted with 20ng of Ela or Elb by a dot-blot assay.

쥐 배종양세포의 Endo-B 유전자 발현

권무식 成均館大學校 科學技術硏究所 1992 論文集 Vol.43 No.2

Endo-B, the mouse form of Keratin 18, is the first member of the large intermediate filament gene family to be expressed during embryogenesis. Thus, the differentiation of cultured murine embryonal carcinoma cells has been used as a convenient model system of early mouse development. F9,22(mouse embryonal carcinoma) cells were treated with retinoic acid (5 x 10_6M/l) for 72 hours, and then they were fixed with the mixture of glutaraldehyde(2%) and formaldehyde(2%) in phosphate buffered saline for 5 min at 4℃. The fixed cells were immunoreacted with the rabbit anti-Endo-B antiserum. Endo-B was visualized by reaction with rhodamine conjugated goat anti-rabbit antibodies. Some of the differentiated F9 cells exhibited strong fluorescence indicating the expression of Endo-B filament.

피르빅산 탈수소 효소의 고체상 면역학적 분석

권무식 成均館大學校 科學技術硏究所 1990 論文集 Vol.41 No.1

A solid-phase electro-blot procedure was used for immunoassay of mammalian pyrurate dehydrogenase(E1) subunits a(Ela) and b(Elb). E1(porcine kidney) was fractionated by sodium-dodecyl sulfate polyacrylamide gel electrophoresis. Ela and Elb proteins were electrophoretically transferred onto nitrocellulose membrane with a constant current of 70mA as a function of time. The adsorbed proteins(Ela & Elb) were immunoreacted with rabbit anti-E1(porcine kidney) antiserum, then the immunoadsorbed Ela and Elb were identified by goat anti-rabbit IgG-horseradish peroxidase conjugate immunoassay kit. It has been found that the solid-phase immunoassay sensitivity of Ela and Elb increased in accordance with increment of transfer time up to a certain point under these experimenal conditions.

Bacillus 殺蟲 蛋白質(CryIC)抗體 生産과 그 應用에 관한 硏究 : Its Application To Plant Biotechnology

권무식 성균관대학교 생명과학자원연구소 1997 生命資源科學硏究 Vol.4 No.2

Bacillus thuringiensis spps., gram-positive soil bacteria, are characterized by their ability to produce crystalline inclusions during sporulation. Inclusions are proteins exhibiting a highly specific entomocidal activity. One of the inclusions, CryIC is specifically toxic to lepidopteran insects. The gene CryIC(7.43kb) has been isolated from Bacillus thuringiensis entomocidus 60.5 and ligated to pTZ vector. The recombinant plasmid named to pSB607. The pSB607 was transformed to E.coli JM109. A number of transformants were selected. The CryIC was purified using differential solubility. The protein was treated with trypsin to increase it's toxicity. They were observed by SDS-PAGE. The activated CryIC was used as an immunogen. Some 600㎍ of the immunogen were hypodermally injected on the hide back & thighs of a rabbit to generate antisera against CryIC. Titration of anti-CryIC antisera has been achieved by blotting or ELISA. In western blot, 2.34ng antigen was detected. A mutant clone of the CrylC is under construction.

Pasteurella multocida의 외막 단백질 H에 의해 유도되는 방어적 항체와 면역

권무식,김영봉,이정민,Kwon, Moo-Sik,Kim, Young-Bong,Lee, Jeong-Min 한국미생물학회 2007 미생물학회지 Vol.43 No.1

Pasteurella multocida is one of the important animal pathogen causing widespread infections in various domestic animals. In swine, it causes severe respiratory diseases such as atrophic rhinitis and pneumonic pasteurellosis. To develop the efficient subunit vaccine against swine atrophic rhinitis, we investigated protective antibodies and humoral immunity of outer membrane protein H (OmpH) which is one of the major outer membrane proteins in P. multocida. Outer membrane fraction of P. multocida was immunologically detectable using antisera from both mice groups vaccinated by formalin-killed whole cells and by commercial vaccine. The expression vector for production of recombinant OmpH was constructed and the recombinant OmpH was expressed and purified from E. coli. Recombinant OmpH showed high antigenic and immunogenic properties in mice vaccination and ELISA with antisera.

Klebsiella pneumonia에서 Quinoprotein의 발현에 대한 면역학적 고찰

권무식 성균관대학교 생명공학연구소 2000 生命工學硏究 Vol.6 No.1

Pyrroloquinoline quinone (PQQ) is an molecule found in many enzymes called quinoproteins, i.e., alcohol dehydrogenases, amine oxidases, decarboxylases, in a broad spectrum of biological materials. It has been considered a very important molecule for physiological property as a enzymatic cofactor. It has been, therefore, interesting to develop a sensitive method to identify the quinoproteins in organisms. Monospecific polyclonal antiserum specific to PQQ was raised in a rabbit as fllow. The PQQ was conjugated to bovine serum albumin (BSA) by 1-Ethyl-3-(3-dimethyl-aminopropyl) carbodiiminde HCI (EDC) mediated reaction. The PQQ-BSA conjugate was purified by gel filtration. The conjugant was emulsified with adjuvant. The emulsion was hyperdermaly injected to a rabbit. PQQ-lipase conjugate was made to use as an antigen for the anti PQQ-BSA conjugate antiserum. The titer of the antiserum was determined by indirect solid phase immunoassay. The antiserum showing the highest titer recognised nano-gram quantity of PQQ-lipase conjugate. Also, we raised anti PQQ antiserum in a rabbit to examine quinoproteins in a prokaryote, Klebsiella, pneumonia.

3-oxoacid CoA-transferase cDNA 클론의 혈청학적 동정

權戊植 성균관대학교 기초과학연구소 1987 論文集 Vol.38 No.2

A cDNA clone for 3-oxoacid CoA-transferase was identified in a λgtll expression gene bank with a monospecific polyclonal rabbit anti-rat 3-oxoacid CoA-transferase. The cDNA clone bank was constructed from poly(A^+) RNA isolated from rat(12 day-old) brains. The cDNA clone was subcloned into M13mp18 and used for further characterization.

Poly(A^+) RNA로 부터 cDNA 의 합성

權戊植 成均館大學校 科學技術硏究所 1987 論文集 Vol.38 No.2

Some datailed procedures for the isolation of biologically active poly(A^+) RNAs from the rat liver and in vitro synthesis of complementary DNA using the isolated poly(A^+) RNA as templates, were described.

미분화된 F9 세포에서 JunB에 의한 AP-1 활동의 유도

權戊植 成均館大學校 科學技術硏究所 1992 論文集 Vol.43 No.1

A full-length JunB cDNA was inserted into an eukaryotic expression vector, pHBAPr-l-neo(or, LK444), to construct a JunB cDNA expression plasmid, LK_4JunB. Undifferentiated murine embryonal carcinoma cells(F9 22 : 5 X 10 exp (6)/10ml) were cotransfected by the calcium phosphate method with DNAs of LK_4LAC(1.5㎍), 3 X TRECAT(3㎍) along with or without PvuII-cut LK_4JunB(8,㎍), in order to test a plausible role of the JunB or AP-1 activity. It was revealed that the cells cotransfected with the LK_4JunB showed some 20 times higher the CAT reporter gene activity than the value obtained from the cells without the LK_4JunB. This finding strongly suggest the fact that the JunB should have the AP-1 activity.

프라스미드 DNA의 대량 분리법 연구 및 아가로스 겔 전기영동상의 고찰

權戊植 成均館大學校 科學技術硏究所 1988 論文集 Vol.39 No.1

A novel method for a large scale isolation of plasmid pBR322 from Escherichia coli HB101 has been developed. The plasmid has been characterized on agarose gel electrophoresis.