• 한국미생물·생명공학회지
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• 제19권5호
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• pp.433-438
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• 1991

Phenylalanine 고생산용 plasmid pSY130-14는 $\lambda$-phage 유래 온도감수성을 가지고 있으므로 $cI_{857}$ repressor와 PL과 PR을 온도를 올려 phenylalanine의 생산을 유도한다. pSY130-14를 갖는 E.coli AT2471은 kanamycin 무첨가, $38.5^{\circ}C$, 48시간에서 약 30%로 plasmid가 없어지며 첨가에서 안정성이 떨어졌다가 배양시간과 더불어 올라갔는데, 이것은 생육에 피요한 kanamycin gene과 ori만이 남는 것으로 생각된다.

• 한국미생물·생명공학회지
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• 제14권4호
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• pp.319-324
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• 1986

Lactobacillus casei 모세포와 동종간 융합세포간의 유산생성, 단백질 분해력, Bacteriophage 저항성 등의 생리적 특성과 표현형을 비교분석하였다. 두 모세포 사이에서 형성된 융합세포는 모세포를 침입하는 bacteriophage에 대하여 저항성을 시험한 결과 L. casei C-M 표현형은 융합세포 중 46%를 차지하였고, L. casei 3-M 표현형은 42%였고, 12%의 융합세포는 두 모세포 bacteriophage에 저항성을 소유하였다. 융합세포의 유산생성력과 단백질 분해력도 유사한 출현율을 나타내었다. Plasmid DNA의 Hind III 분해물은 모세포와 융합세포간에 차이가 없었으나, 융합세포들은 L. casei C-M 또는 L. casei 3-M 중 어느 한 모세포의 Plasmid만을 소유하고 있었다.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.457-463
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• 1999

The physical map of bacteriophage $\PhiFC1$ DNA was constructed with the restriction endonucleases SalI, BamHI, EcoRI, XbaI, and AvaI. The 40.5-kb DNA restriction map is shown to be circularly permuted representing the headful packaging mechanism of the phage. The DNA restriction fragments containing the packaging initiation site(pac) was localized on the restriction map and the nucleotide sequences of the region were analyzed. Four open reading frames (ORFs), following one another with the same orientation, were found at the region. The 2nd ORF (ORF-ts) has significant amino acid sequence homologies to the previously known terminase small subunits of other bacteriophages. The putative terminase small subunit gene has a presumptive NTP-hydrolysis motif and a helix-turn-helix motif. The cleavage site for the first round of packaging was found to be located at the coding sequence of the putative terminase small subunit gene. The fourth ORF, even if partially sequenced, has a good amino acid sequence homology to the portal vertex proteins of other bacteriophages representing the evolutionarily conserved arrangements of genes near the pac site of this bacteriophage, $\PhiFC1$.

• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.385-393
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• 2016

Pseudomonas syringae pv. actinidiae causes bacterial canker disease in kiwifruit. Owing to the prohibition of agricultural antibiotic use in major kiwifruit-cultivating countries, alternative methods need to be developed to manage this disease. Bacteriophages are viruses that specifically infect target bacteria and have recently been reconsidered as potential biological control agents for bacterial pathogens owing to their specificity in terms of host range. In this study, we isolated bacteriophages against P. syringae pv. actinidiae from soils collected from kiwifruit orchards in Korea and selected seven bacteriophages for further characterization based on restriction enzyme digestion patterns of genomic DNA. Among the studied bacteriophages, two belong to the Myoviridae family and three belong to the Podoviridae family, based on morphology observed by transmission electron microscopy. The host range of the selected bacteriophages was confirmed using 18 strains of P. syringae pv. actinidiae, including the Psa2 and Psa3 groups, and some were also effective against other P. syringae pathovars. Lytic activity of the selected bacteriophages was sustained in vitro until 80 h, and their activity remained stable up to 50℃, at pH 11, and under UV-B light. These results indicate that the isolated bacteriophages are specific to P. syringae species and are resistant to various environmental factors, implying their potential use in control of bacterial canker disease in kiwifruits.

• Journal of Microbiology and Biotechnology
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• 제22권6호
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• pp.806-813
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• 2012

Adipocyte differentiation is strongly associated with obesity, which causes metabolic disorders. In this study, we investigated the inhibitory effects of widdrol on 3T3-L1 preadipocyte growth and differentiation. Widdrol decreased lipid droplet accumulation and down-regulated adipogenic transcription factors such as C/$EBP{\alpha}$, C/$EBP{\beta}$, and $PPAR{\gamma}$. Widdrol blocked preadipocyte proliferation and differentiation through the inhibition of mitotic clonal expansion, which was accompanied by the failure of degradation of p21, a cyclin-dependent kinase inhibitor. Cell-cycle analysis clearly indicated that widdrol actively induces cell-cycle arrest at the G1-S phage transition, causing cells to remain in the preadipocyte state. Moreover, widdrol increased p21 expression and inhibited Rb phosphorylation in preadipocyte incubated in a hormone medium. Therefore, these findings clearly suggest that widdrol blocks preadipocyte growth and differentiation through the inhibition of mitotic clonal expansion by p21-and Rb-dependent G1 arrest and can be developed as a potent anti-adipogenic agent for reducing obesity.

• 대한한방부인과학회지
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• 제18권4호
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• pp.10-23
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• 2005

Purpose : This study is to examine the ability of Rhizoma Scirpi (RS) to induce HeLa cell viability. Methods : We culture HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : 1. RS induces mitochondria membrane potential collapse. 2. P38 MAPK is involved in RS-induced death in HeLa cells. 3. P38 MAPK is involved in RS-induced apoptosis in HeLa cells. 4. P38 MAPK reguates RS-induced caspase-3, -8 and -9 activation in HeLa cells. 5. The inhibition of caspase regulates RS-induced cell death in HeLa cells. 6. RS induces mitochondria membrane potential collapse in HeLa cells. 7. P38 MPK is involved in the regulation of Bcl-2 and Bfu in HeLa cells.8. RS regulates the expression of Bcl-2 and Bax in HeLa cells. 9. SR induces p38 MAPK activation in HeLa cells. Conclusion : RS induces apoptosis in HeLa cells via p38 MAPK activation.

• BMB Reports
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• 제37권2호
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• pp.254-259
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• 2004

The wild-type and temperature-sensitive (ts) repressor genes were cloned from the temperate mycobacteriophage L1 and its mutant L1cIts391, respectively. A sequencing analysis revealed that the $131^{st}$ proline residue of the wild-type repressor was changed to leucine in the ts mutant repressor. The 100% identity that was discovered between the two DNA regions of phages L1 and L5, carrying the same sets of genes including their repressor genes, strengthened the speculation that L1 is a minor variant of phage L5 or vice versa. A comparative analysis of the repressor proteins of different mycobacteriophages suggests that the mycobacteriophage-specific repressor proteins constitute a new family of repressors, which were possibly evolved from a common ancestor. Alignment of the mycobacteriophage-specific repressor proteins showed at least 7 blocks (designated I-VII) that carried 3-8 identical amino acid residues. The amino acid residues of blocks V, VI, and some residues downstream to block VI are crucial for the function of the L1 (or L5) repressor. Blocks I and II possibly form the turn and helix 2 regions of the HTH motif of the repressor. Block IV in the L1 repressor is part of the most charged region encompassing amino acid residues 72-92, which flanks the putative N-terminal basic (residues 1-71) and C-terminal acidic (residues 93-183) domains of L1 repressor.

• BMB Reports
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• 제28권6호
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• pp.509-514
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• 1995

A putative secondary structure of the mRNA for the human hepatitis B virus (HBV) X gene is proposed based on not only chemical and enzymatic determination of its single- and double-stranded regions but also selection by the computer program MFOLD for energy minimum conformation under the constraints that the experimentally determined nucleotides were forced or prohibited to base pair. An RNA of 536 nucleotides including the 461-nucleotide HBV X mRNA sequence was synthesized in vitro by the phage T7 RNA polymerase transcription. The thermally renatured transcripts were subjected to chemical modifications with dimethylsulfate and kethoxal and enzymatic hydrolysis with single strand-specific RNase T1 and double strand-specific RNase V1, separately. The sites of modification and cleavage were detected by reverse transcriptase extension of 4 different primers. Many nucleotides could be assigned with high confidence, twenty in double-stranded and thirty-seven in Single-stranded regions. These nucleotides were forced and prohibited, respectively, to base pair in running the recursive RNA folding program MFOLD. The results suggest that 6 different regions (5 within X mRNA) of 14~23 nucleotides are Single-stranded. This putative structure provides a good working model and suggests potential target sites for antisense and ribozyme inhibitors and hybridization probes for the HBV X mRNA.

• 한국추진공학회:학술대회논문집
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• 한국추진공학회 2010년도 제35회 추계학술대회논문집
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• pp.628-635
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• 2010

냉각기계를 적용한 환경조절계통의 성능은 항공기의 비행환경에 따라 엔진의 운전상태가 달라지므로 환경조절장치로 취출되는 공기의 상태(온도와 압력)에 따라 급격히 바뀌게 된다. 즉, 항공기의 운전상태에 따라 환경조절장치는 운용성능을 100%만족하는 영역에서 운전될 수도 있고, 그렇지 않을 수도 있다. 그렇기 때문에, 일반적으로 냉각기계를 적용한 환경조절장치의 설계규격은 항공기에서 요구하는 개발규격보다 큰 설계여유를 갖도록 설계한다. 이로 인한 시스템적인 손실요인을 최소화하기 위하여 본 연구에서는 냉각기계형 환경조절장치에 상변화 물질의 적용가능성을 연구하였다. 그 결과 상변화 현상을 이용하여 환경조절장치의 출구온도를 일정온도 범위 이내로 제어하면, 현재 운용되는 냉각기계형 환경조절장치의 일반적인 성능여유보다 적은 성능여유에서도 환경조절장치에 요구되는 성능요구조건을 만족할 수 있는 것으로 분석되었다. 본 연구 결과 환경조절장치의 운용개념 변경을 통하여 환경조절장치의 시스템 적용성 향상을 기대할 수 있을 것으로 판단된다.

• 자연과학논문집
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• 제14권1호
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• pp.39-50
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• 2004

박테리오 파아지 T7 RNA 중합효소는 다른 RNA 중합효소와 비교하여 볼 때 보조인자 없이 전사를 진행하는 하나의 subunit로 구성된 RNA 중합효소이다. 전사 진행 단계 중에서 T7 RNA 중합효소의 전사연장을 연구하기 위해 biotin이 결합된 DNA 주형을 streptavidin bead로 고정시킴으로서 T7 RNA 중합효소의 진행과정을 관찰할 수 있었고, 이러한 기작을 이용하여 일련의 활성을 가지는 가장 안정한 전사연장복합체들을 얻을 수 있었다. 전사 연장체들은 16번 염기 위치로부터 18번 염기의 위치까지 방사선 동위원소가 표지되어 있으며 이들 표지된 전사연장복합체들은 단계별로 합성하여 22-40개 핵산잔기들이 합성된 전사연장복합체들을 얻을 수 있었다. 이와 같은 전사연장복합체들을 PTH 전사종결 부위가 있는 주형으로 사용하여 야생형 및 R173C 돌연변이 RNA 중합효소를 이용하여 전사연장복합체를 제조하여 비교한 결과 PTH 전사종결에 둔감한 R173C 돌연변이 중합효소의 경우 야생형에 비해 PTH 전사종결부위를 지난 위치에서도 전사연장복합체가 생성되었다.