배추에서 암(暗)조건 발아 유전자의 ESTs 제작

권무식,류종석,김재범,유형진 한국유전학회 1997 Genes & Genomics Vol.19 No.3

Expressed sequence tags (ESTs) are short cDNA sequences generated from randomly selected library clones. ESTs are widely used to clone genes and/or to elucidate their structures and/or functions. We generated a set of ESTs in Chinese cabbage which is an important vegetable species in Korea. Fifty ESTs were generated from the Chinese cabbage (Brassica campestris L. var. pekinensis). Poly A^+ RNAs were isolated from 10-day-old seedlings germinated in dark and two cDNA libraries were constructed by employing λ ZAP-cDNA synthesis system (Stratagene, USA). Then, clones from the libraries were selected randomly and were sequenced by the Sanger method with manual or automated DNA sequencing apparatus. Comparing the ESTs with the coding sequences of the previously reported protein in GenBank or EMBL, significant levels of similarity were found in the amino acids or nucleotide sequences. However, some ESTs showed low level of similarity with the sequences in GenBank or EMBL, which could represent novel genes. Therefore, the reported sequences and cDNA libraries in this study could be utilized to clone more genes in Brassica in the future.

통계학적 학습을 이용한 머리와 어깨선의 위치 찾기

권무식,Kwon, Mu-Sik 한국통신학회 2007 韓國通信學會論文誌 Vol.32 No.2C

영상에서 사람의 머리위치를 찾는 문제에 있어서 어깨선 정보를 이용하는 것은 아주 유용하다. 영상에서 머리 외곽선과 어깨선의 형태는 일정한 변형을 유지하면서 같이 움직이므로 이를 ASM(Active Shape Model) 기법을 사용해서 통계적으로 모델링 할 수 있다. 그러나 ASM 모델은 국부적인 에지나 그래디언트에 의존하므로 배경 에지나 클러터 성분에 민감하다. 한편 AAM(Active Appearance Model) 모델은 텍스쳐 등을 이용하지만, 사람의 피부색, 머리색깔, 옷 색깔 등의 차이로 인해서 통계적인 학습방법을 쓰기가 어렵고, 전체 비디오에서 외모(Appearance)가 시간적으로 변한다. 따라서, 본 논문에서는 외모(Apperance) 모델을 변화에 따라 바꾸는 대신, 영상의 각 화소를 머리, 어깨, 배경으로 구분하는 분별적 외모 모델(discriminative appearance)를 사용한다. 실험을 통해서 제안된 방법이 기존의 기법에 비해서 포즈변화와 가려짐, 조명의 변화 등에 강인함을 보여준다. 또한 제안된 기법은 실시간으로 작동하는 장점 또한 가진다. Associating the shoulder line with head location of the human body is useful in verifying, localizing and tracking persons in an image. Since the head line and the shoulder line, what we call ${\Omega}$-shape, move together in a consistent way within a limited range of deformation, we can build a statistical shape model using Active Shape Model (ASM). However, when the conventional ASM is applied to ${\Omega}$-shape fitting, it is very sensitive to background edges and clutter because it relies only on the local edge or gradient. Even though appearance is a good alternative feature for matching the target object to image, it is difficult to learn the appearance of the ${\Omega}$-shape because of the significant difference between people's skin, hair and clothes, and because appearance does not remain the same throughout the entire video. Therefore, instead of teaming appearance or updating appearance as it changes, we model the discriminative appearance where each pixel is classified into head, torso and background classes, and update the classifier to obtain the appropriate discriminative appearance in the current frame. Accordingly, we make use of two features in fitting ${\Omega}$-shape, edge gradient which is used for localization, and discriminative appearance which contributes to stability of the tracker. The simulation results show that the proposed method is very robust to pose change, occlusion, and illumination change in tracking the head and shoulder line of people. Another advantage is that the proposed method operates in real time.

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.

피루브산 탈수소 효소(송아지 심장)의 항체(토끼)생산

권무식 한국생물공학회 1990 KSBB Journal Vol.5 No.4

Rabbit anti-bovine heart PDH antiserum was raised against El(a, b) isolated from PDC, and then applied to detect Ela and Elb. Appropriate amounis of El were fractionated by SDS-PAGE and electrophoretically transferred to nitrocellulose membrane. The Ela and Elb on the membrane were incubated with anti-El antiserum and identified by GAR-HRP system. It has been found that the immunodetection sensitivity of Ela and Elb were directly proportional to the amount of antigen and transfer time. The lengthy transfer times increased the immunodetection sensitivity of Ela and Elb. The maximal detection sensitivity of Western blotting of Ela and Elb was achieved at 3.5 V/cm for 16-hour transfer under these experimental conditions.

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.

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.

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.

통계학적 학습을 이용한 머리와 어깨선의 위치 찾기

권무식 한국통신학회 2007 韓國通信學會論文誌 Vol.32 No.2

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 multocida는 돼지에서 위축성 비염, 폐렴을 비롯한 다양한 호흡기 질환을 일으키는 병원균이다. 본 연구에서는 돼지 위축성 비염에 대한 효과적인 순수정제 백신을 개발하고자 하는 기초 연구로서 P. multocida의 외막 단백질 H에 의해 유도되는 방어적 항체와 면역을 확인하였다. P. multocida의 외막 단백질을 포함하는 분획은 호흡기 질병 혼합 백신에 대한 항혈청과 불활화된 사균 세포에 대한 항혈청 모두에서 면역학적으로 검출 가능하였다. 선행 연구에서 분리한 외막 단백질 H 유전자는 재조합 발현 백터 제작에 이용되어 대장균으로부터 재조합 외막단백질 H를 정제하였다. 실험 동물 면역과 항혈청의 교차반응, ELISA를 통한 항체 역가의 측정 및 공격접종을 통하여, 재조합 외막 단백질 H는 높은항원성을 가지며, 지속적인 체액성 면역을 유도하는 것을 확인하였다. 외막 단백질 H는 순수정제 항원으로서 P. multocida에 의한 호흡기 질환에 대한 효과적인 방어를 유도할 수 있는 단위 백신 후보 단백질로 여겨진다. 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.