• Korean Journal of Microbiology
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• v.28 no.2
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• pp.91-97
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• 1990

Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.

• Biomolecules & Therapeutics
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• v.18 no.1
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• pp.1-15
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• 2010

Progress in the development of DNA vaccines and their delivery strategies has been made since their initial concept as a next generation vaccine. Since DNA vaccine includes non-infectious DNA parts of pathogens, it can't cause disease yet it closely mimic the natural process of infection and immune responses. Despite their early promising results of controlling infectious diseases and cancer in small animal models, DNA vaccines failed to display a level of immunogenicity required for combating these diseases in humans, possibly due to their lower protein expression levels. However, increasing evidence has shown that DNA vaccines are clinically well-tolerated and safe. Furthermore, one notable advantage of DNA vaccines includes convenient utilities of plasmid DNAs coding for antigens. For instance, any emerging pathogens could be prevented easily and timely by allowing the simple exchange of antigen-encoding genes. In this review, newly developed DNA vaccine strategies, including electroporation, which has emerged as a potent method for DNA delivery, targeting infectious diseases and cancer will be discussed with a focus on any on-going DNA vaccine trials or progress made pre-clinically and in clinics.

• Proceedings of the KIEE Conference
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• 1999.11c
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• pp.576-578
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• 1999

본 논문에서는 생물학적인 DNA와 유전자 알고리즘의 진화 메커니즘에 근거를 둔 DNA 코딩방법을 변형하여 새로운 DNA 코딩 방법을 제안한다. 이 방법은 기존의 DNA 코딩 방법이 DNA 유전자의 Redundancy와 Over-lapping 성질 때문에 갖고 있는 DNA 자체의 특성인 염색체의 길이를 자유자재로 변화시킬 수 있는 코딩 기술에 진화단계에서 변형을 가할 수 있는 새로운 유전자 알고리즘을 추가하여, 초기에 국소해로 접근하는 일반적인 유전자 알고리즘의 위험 부담률을 줄이고, 전역 해로의 접근 가능성을 높이는 방법을 제시한다. 또한. 이 변형된 DNA 코딩 방법의 가능성을 입증하기 위하여 시스템 제어에 필요한 지식을 표현하는 적당한 퍼지 규칙을 후건부의 매개변수의 동조만을 통하여 획득하고, 이 규칙에 변형된 DNA 코딩 방법을 적용하여 최적화 된 새로운 퍼지규칙 획득 알고리즘을 개발한다. 제안된 알고리즘을 이용한 퍼지 제어기를 설계하고. 이 제어기의 유용성을 입증하기 위하여 병렬형 이중 도립진자 시스템에 적용하여 시뮬레이션을 실행한 결과 효과적으로 퍼지규칙을 획득하고 제어함을 알 수 있다.

• Korean Journal of Plant Resources
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• v.23 no.6
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• pp.526-534
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• 2010

Genomes of clusters of related eukaryotes are now being sequenced at an increasing rate. In this paper, we developed an accurate, low-cost method for annotation of gene prediction and exon-intron structure. The gene prediction was adapted for delta 1-pyrroline-5-carboxylate-synthetase (p5cs) gene from China wild-type of the halophytic Leymus chinensis (Trin.), naturally adapted to highly-alkali soils. Due to complex adaptive mechanisms in halophytes, more attentions are being paid on the regulatory elements of stress adaptation in halophytes. P5CS encodes delta 1-pyrroline-5-carboxylate-synthetase, a key regulatory enzyme involved in the biosynthesis of proline, that has direct correlation with proline accumulation in vivo and positive relationship with stress tolerance. Using analysis of reverse transcription-polymerase chain reaction (RT-PCR) and PCR, and direct sequencing, 1076 base pairs (bp) of cDNA in length and 2396 bp of genomic DNA in length were obtained from direct sequencing results. Through gene prediction and exon-intron structure verification, the full-length of cDNA sequence was divided into eight parts, with seven parts of intron insertion. The average lengths of determinated coding regions and non-coding regions were 154.17 bp and 188.57 bp, respectively. Nearly all splice sites displayed GT as the donor sites at the 5' end of intron region, and 71.43% displayed AG as the acceptor sites at the 3' end of intron region. We conclude that this method is a cost-effective way for obtaining an experimentally verified genome annotation.

• Journal of the Korean Institute of Intelligent Systems
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• v.9 no.4
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• pp.389-395
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• 1999

최근 몇 년간 생물학적 발생에 대한 구조 및 둥작원리의 모델링에 대한 빠른 진전이 일어나고 있다. 세포자동자(cellular automata CA)와 린드마이어-시스템(L-system)은 다세포의 대표적인 발생/발달 모델이다. L-시스템은 식물의 그래픽 표현에 적용되어 오고 있으며 CA는 인고생명의 연구모델과 인공두뇌의 건축 등의 분야에 적용되어 오고 있다, 현재까지 CA와 L-시스템의 발생규칙은 설계자의 설계에 의존하고 있다. 그러나 진화연사방법을 도입하면 CA와 L-시스템을 자동으로 설계할수 있다. 발생규칙의 진화를 위해서는염색체의 코트화가 필요하다. DNA 코딩방법은 유전자의 중복과 여분을 가지고 있으며 규칙의 표현에 적합한 코딩방법이다. 본 논문에서는 CA와 L-시스템의 규칙을 진화시키기 위한 DNA 코딩 방법을 제안한다.

• Journal of Korean Society of Forest Science
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• v.87 no.3
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• pp.334-340
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• 1998

This research was conducted to identify plant regulatory regions by gene tagging method. A promoterless GUS coding sequence was introduced to Populus nigra ${\times}$ P. maximowiczii via Agrobacterium strains(LBA4404/EHA101), and putative transgenic poplars were selected by culturing on medium containing G418($60mg/{\ell}$) and by GUS assay. Among them one positive plant was to amplify the native sequences flanking to the introduced GUS gene in plant genome by inverse PCR method and from this 730 by DNA product was obtained. After subcloning and sequencing, it has 88% homology to the Eucalyptus gunnii CAD(cinnamyl alcohol dehydrogenase) gene. The GUS gene fused with the putative promoter reinserted into poplar leaves by particle bombardment method to test the funtional promoter activity. Upon staining with X-gluc, many blue spots appeared on the leaf segments bombarded by the chimeric gene 2-3 days, thus the isolated DNA fragment contain some possible coding region as well as a putative regulatory sequences of poplar CAD gene.

• Proceedings of the Korea Information Processing Society Conference
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• 2002.11a
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• pp.319-322
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• 2002

현재 에이전트는 강화 학습 모델을 토대로 사용자의 간섭 없이 사용자 의도를 파악하며 능동적으로 행동하는 기술들이 발달되어 왔다. 하지만 인터넷을 기반으로 한 계획이나 학습 등을 위하여 보다 지적인 능력을 갖춘 에이전트의 기술이 요구된다. 따라서 본 논문에서는 DNA 코딩 기법을 이용하여 사용자의 프로파일을 학습하고. 사용자를 분류하는 AUA(Agent for learning Users' Action)를 제안하고자 한다. AUA는 사용자 학습 에이전트로 사용자의 행위를 관찰하고 행위서열을 생성하고 구분함으로써, 사용자의 관심정도를 보다 세밀하게 분석하고 계획할 수 있다. 또한 AUA는 에이전트간에 관계를 설정함으로 사용자에게 보다 나은 정보 검색을 지원할 수 있다.

• Tuberculosis and Respiratory Diseases
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• v.39 no.3
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• pp.242-247
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• 1992

Background: The ability to precisely measure specific mRNA levels by hybridization to complementary DNA probes is an important tool for analyzing the regulation of gene expression. Surfactant proteins have important roles in regulating surfactant metabolism as well as in determing its physical properties. Method: The complete coding regions for rat surfactant protein complementary DNA of surfactant protein B were subcloned into pGem 3Z or 4Z such that either antisense or sense transcripts were obtained by using SP 6 RNA polymerase. Surfactant protein B mRNA was measured by filter hybridization. Results: Equation of standard curve between counts per minute (Y) and surfactant protein B mRNA transcript input (X) was Y=2034.9 X+159.1. Correlation coefficient was 1.0. Couclusions: Filter hybridization assay is suited to situation when rapid, accurate quantitation of multiple samples is required.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.17
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• pp.7935-7941
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• 2015

The aim of this study was to implement massive parallel sequencing (MPS) technology in clinical genetics testing. We developed and tested an amplicon-based method for resequencing the BRCA1 and BRCA2 genes on an Illumina MiSeq to identify disease-causing mutations in patients with hereditary breast or ovarian cancer (HBOC). The coding regions of BRCA1 and BRCA2 were resequenced in 96 HBOC patient DNA samples obtained from different sample types: peripheral blood leukocytes, whole blood drops dried on paper, and buccal wash epithelia. A total of 16 random DNA samples were characterized using standard Sanger sequencing and applied to optimize the variant calling process and evaluate the accuracy of the MPS-method. The best bioinformatics workflow included the filtration of variants using GATK with the following cut-offs: variant frequency >14%, coverage ($>25{\times}$) and presence in both the forward and reverse reads. The MPS method had 100% sensitivity and 94.4% specificity. Similar accuracy levels were achieved for DNA obtained from the different sample types. The workflow presented herein requires low amounts of DNA samples (170 ng) and is cost-effective due to the elimination of DNA and PCR product normalization steps.

• Molecules and Cells
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• v.22 no.2
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• pp.175-181
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• 2006

Delivery of DNA vaccines to airway mucosa would be an ideal method for mucosal immunization. However, there have been few reports of a suitable gene delivery system. In this study we used a cationic emulsion to immunize mice via the intranasal route with pCMV-S coding for Hepatitis B virus surface antigen (HBsAg). Complexing pCMV-S with a cationic emulsion dramatically enhanced HBsAg expression in both nasal tissue and lung, and was associated with increases in the levels of HBs-specific Abs in serum and mucosal fluids, of cytotoxic T lymphocytes (CTL) in the spleen and cervical and iliac lymph nodes, and of delayed-type hypersensitivity (DTH) against HBsAg. In contrast, very weak humoral and cellular immunities were observed following immunization with naked DNA. In support of these observations, a higher proliferative response of spleenocytes was detected in the group immunized with the emulsion/pCMV-S complex than in the group immunized with naked pCMV-S. These findings may facilitate development of an emulsion-mediated gene vaccination technique for use against intracellular pathogens that invade mucosal surfaces.