• Korean Journal of Microbiology
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• v.51 no.1
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• pp.1-6
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• 2015

The term "P2 sir-associated helper inefficiency" has been used to define the inefficient helper capability of P2 sir mutants for their satellite bacteriophage P4. The aim of this study was to investigate the factors overcoming P2 sir-associated helper inefficiency. At first, we verified whether the P2 cos region containing P4 sid71 cosP2 could overcome P2 sir-associated helper inefficiency with P2 sir3. The result was that P4 sid71 cosP2 could not overcome P2 sir-associated helper inefficiency with P2 sir3. Instead of cos region of P2, the size of the DNA packaged into a $P2_{sir}$-sized head seems to be important for overcoming P2 sir-associated helper inefficiency. In the present work, three kinds of P4 derivatives with packaged DNA sizes between those of P4 ost1 and P4 ost2, were constructed through DNA manipulation. In one P4 derivative, P4 sid71 delRI::apr, the size of the packaged DNA was identified with a CsCl buoyant equilibrium density gradient experiment. According to the burst sizes of the P4 derivatives, they could overcome P2 sir3-associated helper inefficiency. The size of the P4 derivative DNA suitable for packaging into a $P2_{sir3}$-sized head was 28-29 kb.

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.483-490
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• 1992

In order to understand the transfer behavior of a particular gene in water environments, kanamycin resistance ($Km^r$) gene was tracked by Southern hybridization with DNA probe in its conjugal transfer. A $Km^r$ natural bacterial isolate and genetically modified microorganisms (GMMs) constructed from the isolate were used as donor for conjugal transfer of the $Km^r$ gene. The transfer frequencies of the $Km^r$ gene from GMM strains were generally 10 to 100 times higher than those from the natural isolate. The conjugants obtained from GMM strains in LB broth had more plasmids newly appeared, and particularly the conjugants in A Wand FW waters revealed more rearrangement in their plasmids as a function of conjugation time. When plasmids of the conjugants obtained in LB broth were Southern hybridized with DNA probe of the $Km^r$ gene, the $Km^r$ plasmids in the conjugants were detected at the same position of the plasmids in donor cells, in spite of the fact that the plasmids were highly rearranged in conjugant cells. But the $Km^r$ plasmids in the donor of DKI and DKC601, and DKC600 were not identified in the conjugants obtained after 50 h conjugation in AW and after 30 h in AW, respectively. The size of the $Km^r$ plasmids showing hybridization signal were a little changed in the conjugants obtained in A Wand FW waters. Therefore, the method of Southern hybridization with DNA probe was proved to be very specific and useful for tracking of particular genes in water environments.

• KSBB Journal
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• v.11 no.2
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• pp.193-200
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• 1996

In molecular biology, type-II restriction endonuclease, which specifically recognize and cleave DNA at a limited number of sites, have been exploited as a means of characterizing DNA fragments, DNA mapping for genetic engineering. Type-II restriction endonucleases have been found to modulate their substrate specificity under modified conditions such as extreme pH, ionic strength, high enzyme concentration, substitution of metallic cofactors or addition of organic solvents. This study was initiated to investigate the modification of recognition specificity of BamHI according to the different pH and organic solvent under the given buffer condition. The specificity of BamHI is highly depends on the presence of hydrophobicity (LogP: partition coefficient) and pH of reaction solution. The specificity of BamHI is changed in range of LogP -1.03∼-1.35(at pH 7.5), -1.03∼-2.5 (at pH 8.0), -0.75∼-0.25(at pH 8.5), 0.32∼-2.5(at pH 8.9), respectively. Alteration of specificity appears in lower concentration of organic solvent when the reaction occurs in more alkali pH. For example, in DMSO solution, alteration of specificity appears in 20% concentration at pH 7.5 but in 4% concentration at pH 8.9.

• Journal of Life Science
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• v.14 no.1
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• pp.110-114
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• 2004

The optimized RAPD-PCR conditions, that can be utilized as a basic information for analysis of the gelletic characteristics were developed for genetic analysis of four mulberry varieties, named Milsung, Chungil, Suil, and Hansung using a primer, OPY15 (5'-AGTCGCCCTT-3') from Operon company. We tested several different factors for best PCR condition including concentrations of DNA, primer, Mgclu annealing temperature, number of PCR cycle, and prosence/absence of pre-heating time at the begining of PCR reaction in the $25 \mul$volume. The best RAPD profiles were obtained using 50 ng of DNA, 1 $\mu$M of primer, $1 \mum$of $MgCl_2\;,45^{\circ}C$ of annealing temperature and an absence of pre-heating time. An establishment of the stable and reproducible RAPD-PCR conditions are expected to be useful for the subsequent RAPD-related investigation, such as genetic characterization of the mulberry varieties, re-establishment of phylogenetic relationships and development of new varieties.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.39-45
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• 2000

Two maize transposable elements, immobilized Ac (iAc) and Ds, have been introduced into the genome of a diploid potato clone (Solanum tuberosum Group Phureja clone 1.22). The iAc is a modified Ac that is supposed to be unable to transpose but is expected to trans-activate the transposition of a Ds that is unable to transpose by itself. When the leaf and stem explants of in vitro shoots of the clone 1.22 were inoculated with Agrobacterium tumefaciens strains harboring binary vectors containing the iAc and the Ds, calli were formed from the explants on media containing 50 mg/L of kanamycin, and shoots were regenerated from the calli. The regenerated shoots formed roots when cultured on media containing 100 mg/L of kanamycin, whereas untransformed shoots did not form roots on the same media. The PCR amplification of the DNA's from the transgenic plants confirmed that the iAc and the Ds elements were introduced into the potato genome of 1.22.

• Journal of Life Science
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• v.2 no.2
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• pp.120-131
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• 1992

효소에 의한 펩티드합성법은 합성반응의 개발초기에는 용매로서 침전계를 이용하여 아미노산 혹은 펩티드기질에서 펩티드생성의 방향으로 평형화하는 방법을 많이 이용하였지만 최근에는 물에 섞이는 유기용매들을 수용액과 함께 사용하여도 합성반응이 이루어진다는 사실이 밝혀졌다. 하지만, 효소를 공업적으로 이용할 때 값이 너무 비싸고 합성물과의 분리, 정제에 어려움이 있다. 이러한 문제를 해결하기 위하여 효소의 고정화법이 널리 이용되고 있다. 한편 protease에 의한 펩티드의 역합성은 단백질공학의 한 수법인 반합성으로서 주목을 끌고 있다. DNA의 조작기술이 오늘날과 같이 발전하기 이전에는 단백질합성의 공학적기법은 이 반합성과 화학적수식으로 한정되었다. 본고에서는 protease를 이용한 생리활성펩티드의 합성에 필요한 protease의 종류, 이들의 펩티드합성작용기구, 합성설계조건 및 유용 생리활성펩티드의 합성 예를 소개하였다.

• KOREAN POULTRY JOURNAL
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• v.33 no.3 s.377
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• pp.104-106
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• 2001

작년 6월에 인간 유전자 지도의 초안이 작성된 이후 지난 2월 11일에 신문과 방송은 사람이 게놈지도가 완성되었다고 주요뉴스로 보도하였다. 1990년 8월에 인간게놈 연구계획이 발표된지 11년여간의 연구 끝에 이룩한 연구결과이다. 최근, 학문의 급속한 발전으로 질병의 진단과 치료에 새로운 지평이 열리고 있다 수의분야에서도 예외는 아니다. 특히 질병예방에 있어 중요한 백신의 연구개발에 있어서도 괄목할 만한 발전이 있었다. 유전공학의 발전과 여러 병원체의 병원성 인자와 체내의 면역기능과의 관련성 연구결과로 여러질병이 백신으로 예방이 가능하게 되었으며 보다 안전하고 효과적인 새로운 개념의 백신도 개발되고 있다. 새로운 백신, 즉 신백신은 유전자 조작(재조합)에 의한 병원성이 약해진 백신에서부터 여러 질병을 동시에 예방하려고 공안된 벡터백신, 그리고 유전자를 접종하는 DNA 백신 등이 대표적이다. 이들 백신은 기존백신에 사용되고 있는 백신보다 안전하고 경제적이며 우수한 효능을 발휘하도록 고안되고 있다.

• Annual Conference of KIPS
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• 2006.11a
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• pp.69-72
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• 2006

최근 생명 정보학 기술의 발달로 마이크로 단위의 실험조작이 가능해짐에 따라 하나의 chip상에서 전체 genome의 expression pattern을 관찰할 수 있게 되었고, 동시에 수 만개의 유전자들 간의 상호작용도 연구가능하게 되었다. 이처럼 DNA 마이크로어레이 기술은 복잡한 생물체를 이해하는 새로운 방향을 제시해주게 되었다. 따라서 이러한 기술을 통해 얻어진 대량의 유전자 정보들을 효과적으로 분석하는 방법이 시급하다. 본 논문에서는 마이크로어레이 실험에서 다양한 원인에 의해 발생하는 잡음(noise)을 줄이거나 제거하는 과정인 정규화과정을 거쳐 특징 추출방법인 SVM(Support Vector Machine) 방법을 이용하여 데이터를 2개의 클래스로 나누고, 표준화 방법들의 성능 비교를 위해 Adaptive Simulated Annealing 알고리즘으로 정확도를 평가하는 분류 시스템을 설계 구현하였다.

• Proceedings of the Korean Information Science Society Conference
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• 2001.10a
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• pp.280-282
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• 2001

분자생물학의 급진적 발전은 현대 계통분류학에 큰 변혁을 가져왔다. 특히 유전의 근원물질인 DNA나 RNA를 분리.조작.분석하는 기술의 발전으로 이를 이용만 계통수 제작은 계통생물학의 중요한 실험방법으로 자리잡고 있다. 그 중 염기서열 비교 방법은 현재 유전자 계통수 제작에 가장 널리 이용되는 방법이다. 하지만 이러만 계통수는 각 객체간의 거리만을 표현하고, 객체군간의 차이는 설명하기 힘들다. 본 연구에서는 염기서열의 상대적인 특징(유사도)을 대신하는 염기서열의 총량과 염기 함량 등을 이용해 새로이 분류 기법 중 결정트리 방법에 적응하고, 종 분류의 유전적 모델을 설계한다. 또한 결정트리의 클래스인 종은 상위 클래스들을 포함하고 있어, 본 논문에서는 기존의 결정트리 분류자를 수정한 단계적 결정트기 분류자를 제안한다.

• Tuberculosis and Respiratory Diseases
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• v.43 no.1
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• pp.30-37
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• 1996

Background: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. Methods: We used the $InstaGene^{TM}$ DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1,245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2,536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. Results: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value. Conclusion: Even though both methods had lower possibility of cross contamination, shorter time requirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usefulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.