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.

웨스턴 전이 : 피루브산 탄산효소 (흰쥐 간) Pyruvate Carboxylase (Rat liver)

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

Pyruvate carboxylase isolated from the mammalian rat and mouse livers were subjected to sodiumdodecyl sulfate-polyacrylamide gel electrophoresis. The pyruavte carboxylase on the gel was electrically transferred to pure nitrocellulose membrane (average pore size : 0.2㎛). The enzyme (antigen) immobilized on the nitrocellulose membrane, was identified by a double antibody assy. The Bio-Rad ImmunBlot(Goat Anti Rabbit-Horse Radish Peroxidase Conjugate) Assay Kit (Bio Rad Co.) was employed here. The electroblot conditions were best optimized for the pyruvate carboxylase. The detailed procedures were discussed in the text.

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.

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.

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.

한국인 혈액응고 제 9인자의 제한효소 절편 길이 다형성 분석

권무식(Moo Sik Kwon),이정민(Jeong Min Lee),전봉균(Bong Kyun Jeon),오성관(Sung Gwan Oh),류종석(Chong Suk Ryou),오보훈(Bo Hoon Oh) 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.3

N/A Objective : The purpose of this study was to investigate the methods for analysis of restriction fragment length polymorphisms of hemophilia B (coagulation factor Ⅸ) gene in Korean population. Methods : Genomic DNAs were extracted from 40 Korean females. In order to amplify genomic DNAs at the region of the polymorphic sites, two sets of primers (Hha I and Dde I) were synthesized. The primers were named as FIX1, FIX2 for Hha I, and Dde I 59, Dde I 39 for Dde I, respectively. Hha I primers annealed 3'-flanking region of the Factor Ⅸ gene and amplified 230 bp long fragment. The PCR fragment (230 bp) treated with Hha I endonuclease produced two fragments (150 bp and 80 bp), when the polymorphic site existed. Dde I primers annealed the region of the first intron of Factor Ⅸ gene and amplified 319 bp long fragments. People cases with Dde I polymorphic site are supposed to produce 369 bp long fragment. Results : It has been found that seven (14 X chromosomes) out of forty individuals showed Hha I polymorphism. However, none of the experimental People cases showed the Dde I polymorphism. Conclusion : By the analysis of 80 chromosomes, the PICs calculated from allele frequency of Hha I-RFLP (0.175/0.825) and that of Dde I-RFLP (0.0/1.0) were 0.289=[1-(0.1752+0.8252)] and 0=[1-(02+12)], respectively. From these results, it can be postulated that Hha I and Dde I polymorphisms of the Factor Ⅸ gene in Korean exhibited different patterns from those of Caucasian.

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.

통계학적 학습을 이용한 머리와 어깨선의 위치 찾기

권무식 한국통신학회 2007 韓國通信學會論文誌 Vol.32 No.2

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.