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특정 염기변환 방법을 이용한 PU.1 유전자의 돌연변이체 제조
류종석,권무식 成均館大學校 科學技術硏究所 1992 論文集 Vol.43 No.2
The purine-rich sequence (GAGGAA) binding protein, PU.1, is one of the tissue specific transcription factors. It is related to the activation of macrophage & B-cell in mammals. It is postulated that the activation of PU.1 could be accomplished by the phosphorylation of serine(s). The PU.1 contains 22 serine residues in its coding region. In order to figure out a plausible phosphorylation site, one nucleotide Thymine of PU.1 cDNA coding the 126th amino acid serine (TCC) was substituted with Guanine by site-directed mutagenesis using polymerase chain reaction. The mutated PU.1 cDNA was ligated into a pBluescript KS+ and transformed into E. coli JM109. The mutant clone will be employed to elucidate the phosphorylation site(s) and to understand the phosphorylation mechanism of transcription factor, PU.1.
라즈베리파이에서 MIRACL을 활용한 암호함수 연산시간 분석
류종석(Ryu Jong Seok),권덕규(Kwon Deok Kyu),손승환(Son Seung Hwan),이준영(Lee Joon Young),박영호(Park Young Ho) 한국통신학회 2021 한국통신학회 학술대회논문집 Vol.2021 No.6
모바일 디바이스의 보급 확산으로 IoT(Internet of Things) 환경은 스마트 홈, 스마트 시티 및 헬스 케어 등 다양한 분야에 사용되고 있으며 제한된 환경에서 사용자에게 안전한 서비스를 제공하기 위한 효율적인 보안 프로토콜 연구가 활발히 진행되고 있다. 본 논문에서는 프로토콜 성능 분석을 위해 MIRACL(Multiprecision Integer and Rational Arithmetic Cryptography Library)을 활용하여 Sutrala 등과 Das 등이 제안한 프로토콜에서 사용된 암호함수 연산시간을 분석한다.
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.
습식 합성법에 의한 고순도 ${\alpha}-Al_2O_3$ 미세분말의 합성 연구
최진호,류종석,한양수,김준,이현국,김혁년,Jin-Ho Choy,Jong-Seok Yoo,Yang-Su Han,Joon Kim,Hyeon-Kook Lee,Hyuk-Nyun Kim 대한화학회 1991 대한화학회지 Vol.35 No.3
고순도 암모니움 명반 결정을 합성한 후 열분해 시켜 높은 순도의 초미세 산화알루미늄(${\alpha}-Al_2O_3$)분말을 합성하였다. 이 때 불순물인 Na$_2$O의 혼입과 Al(OH)$_3$의 침전을 최대한으로 방지하기 위해 pH = 1.5∼2.5의 영역에서 암모니움 명반을 합성하였으며, pH 조건은 수용액 중에서 Na와 Al 이온의 수산화물과 탄산염 형성을 고려, pH에 따른 각 이온종들의 농도가 이론적으로 계산되었다. 그 결과 ${\alpha}-Al_2O_3$의 순도는 99.7%이상이고, 입자는 ${\phi}$ = 0.1∼0.5 ${\mu}$m의 균일한 크기의 분말이 얻어졌다. Ultra-fine alumina, ${\alpha}-Al_2O_3$, with ${\phi}$ = 0.1∼0.5 ${\mu}$m was obtained from pure ammonium aluminum sulfate(alum) as the thermal decomposition product. Pure alum(> 99.7%) could be prepared by the precepitation and the successive recrystallization in an acidic aqueous solution at pH = 1.5∼2.5, which was theoretically predicted by only considering the concentrations of hydroxide and carbonate for aluminum and sodium in the solution, and also experimentally confirmed as the optimum precepitation condition for alum without forming any impurities like aluminum hydroxide or sodium one.
장시혁,류종석,김재범,이풍연,권무식 성균관대학교 생명과학자원연구소 1996 生命資源科學硏究 Vol.3 No.2
PU.1 is a transcription factor which binds to a DNA motif, 5'-GAGGAA-3' (PU Box). Its expression is restricted to macrophage and B cell in mammal. Activation of the PU.1 transcription factor seems to be accomplished by the phosphorylation of specific serine residue(s) of PU.1 the polypeptide. In an attempt to elucidate the phosphorylation site(s), a PU1 mutant clone named pJKS142A was constructed as follows. Two nucleotides coding for the 142th amino acid [serine(AGC)] of the PU1 polypeptide, "Adenine & Guanine", were substituted by "Guanine & Cytosine" by employing splicing by overlap extenion (SOE) methods. The mutant PU.1 cDNA, PU.S142A, was ligated into HindⅢ/XbaI-cut pBluescript KS+, and then transformed into E. coli XL-1 Blue to generate pJKS142A. Recombinant plasmid DNA, pJKS142A was isolated and the DNA sequence was determined by the method of Sanger to confirm the site directed mutagenesis of PU.1 eDNA. The pJKS142A will be instrumental in evaluating a plusible phosphorylation site(s) of PU.1.
Bacillus thuringiensis 살충성 결정단백질 유전자(cry II A)의 형질전환 식물 제작
이정민,류종석,권무식 한국식물생명공학회 1997 식물생명공학회지 Vol.24 No.5
Bacillus thuringiensis는 그람 양성 토양 세균으로 포자형성시 결정화된 내포체를 형성하는데, 이 내포체를 구성하는 결정단백질은 각 곤충에 대하여 특이적인 독성을 나타낸다. 살충성 결정단백질 중 Cry II A 결정단백질은 인시류와 쌍시류 곤충에 모두 특이적으로 작용한다. 결정단백질은 살충제로서 불안정하고 포장에서 지속성이 낮은 단점을 가지고 있으므로, 본 연구에서는 Cry II A 결정단백질 유전자가 형질전환된 담배 식물을 제작하고자 하였다. cry IIA 유전자가 삽입된 벡터를 대장균에 형질전환한 후, 알칼리 용액에 대한 용해도의 차이를 이용하여 Cry II A 결정단백질(70 kDa)을 분리하였고, 분리한 Cry II A 결정단백질을 trypsin 처리하여 활성화된 Cry II A (50 kDa)를 확인하였다. 식물 형질전환을 위하여 두 개의 CaMV 35S promoters에 의해 발현이 조절되는 식물 발현 벡터에 cry II A 유전자를 클로닝 하였다. 이 식물 발현 벡터를 Agrobacterium을 이용한 엽편형질 전환을 통해 담배(N. tabacum var. Petit Havana SRI)에 형질전환 시켰으며, 재분화 과정을 거쳐 여섯 개체의 형질전환 식물체를 얻었다. Southern blot을 통하여 분석한 결과 세 개체 내에 cry II A 유전자가 존재하였는데, 한 개체에는 하나의 cry II A 유전자가, 또 한 개체에는 두 개의 cry II A 유전자가, 다른 한 개체에는 잘려진 형태의 cry II A 유전자가 존재함을 확인할 수 있었다. 본 연구를 통하여 얻어진 cry II A 형질전환 식물체는 살충성 검정 등을 거친 후 내충성 식물 생산 및 후대 유전 양상 분석을 위한 기초 자료로 이용될 수 있을 것이다. Bacillus thuringiensis, a gram-positive soil bacterium, is characterized by its ability to produce crystalline inclusions during sporulation. The crystal proteins exhibit a highly specific insecticidal activity. An insecticidal crystal protein (ICP), Cry II A, is specifically toxic to both lepidopteran and dipteran insects. In this study, tobacco plants transformed by the cry II A gene have been generated. The Cry II A crystal protein was purified from E. coli JM103 harboring cry II A gene by differential solubility. The activated Cry II A was prepared by tryptic digestion. The purified protoxin (70 kDa) and the activated toxin (50 kDa) were analyzed by SDS-PAGE. To generate the transgenic tobacco having cry II A gene, the cry II A gene was subcloned to a plant expression vector, pSRL2, having two CaMV 35S promoters. The recombinant plasmid was transformed into tobacco (N. tabacum var. Petit Havana SR1) by Agrobacterium-mediated leaf disc transformation. Through the regeneration, six putative transgenic tobacco plants were obtained and three transformants were confirmed by Southern blot analysis. It has been found that one plant had single copy of cry II A gene, another had two copies of the gene, and the third had a truncated gene. After the immunochemical confirmation of cry II A expression in plants, the transgenic tobacco plants will be used to study the genetics of future generation with the insecticidal crystal protein gene cry II A.