• 전자공학회논문지SC
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• 제48권2호
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• pp.86-90
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• 2011

전계효과 트랜지스터(FETs)를 이용한 전하 검출형 DNA센서는 DNA가 가지고 있는 음전하를 중성화 시키는 양이온의 영향은 매우 중요하다. 본 논문에서는 양이온 농도에 의존하는 Debye length에 관한 연구를 통해 DNA 검출감도를 평가하였다. Debye length는 낮은 농도의 NaCl 용액에서 긴 거리를 유지하며, Debye length가 높은 용액에서 DNA가 가지고 있은 음전하는 게이트 채널에 보다 많은 영향을 미친다. 용액내 NaCl농도가 1 mM인 버퍼 용액에서 상보적 DNA의 hybridization에 의한 전계효과 트랜지스터의 게이전압은 21 mV 시프트 했으며, NaCl 농도가 10 mM인 버퍼 용액에서는 7.2 mV, NaCl농도가 100 mM인 버퍼 용액에서는 전계효과 트랜지스터의 게이트 전압이 5.1 mV 각각 시프트 하였다. 이러한 결과를 바탕으로 전계효과 트랜지스터를 이용한 전하 검출형 DNA센서의 검출 감도는 Debye length에 의존하는 것을 규명하였다.

• 약학회지
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• 제31권2호
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• pp.116-125
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• 1987

It is still difficult and time consuming to obtain cDNA sequences that contain the entire nucleotide sequence of the corresponding mRNA. A rapid and high efficient cloning method to obtain full-length cDNA segments is thus developed. The cloning procedure described here consists of the construction of oligo(dT)-tailed vector primer using pWR34 plasmid, polyadenylation of mRNA-cDNA heteroduplex using terminal deoxytransferase, and replacement of MRNA strand with DNA by RNase H and DNA polymerase I. The restriction endonuclease analysis shows that the size of inserted-cDNA is in the range of 1.5~4.0 kb long suggesting that most of cloned cDNA are full-length or nearly full-length cDNA. The plasmid-DNA recombinants obtained were 4$\times$$10^5$~$10^{6}$ per $\mu\textrm{g}$ of rat liver poly (A$^+$)mRNA, which is 4 to 10 fold higher cloning efficiency in comparison to the presently used methods for full-length cDNA cloning. The results indicate that the described cloning system is much simpler, less time consuming, and very efficient cloning method to construct a cDNA library.

• The Plant Pathology Journal
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• 제18권4호
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• pp.187-191
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• 2002

This paper describes the cloning and sequence analysis of the 5'-terminal region and full-length cDNA production of genomic RNA of Lily symptomless virus (LSV), a Species Of the genus Carlavirus. A sing1e DNA band about 600 bp harboring the 5'-end of genomic RNA of the virus was successfully amplified by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), and was cloned for nucleotide sequence determination. Sequence analysis of selected RACE cDNA clones revealed that the LSV 5'non-translated region consists of 67 nucleotides long of AT rich stretch followed GC rich from the 5'-end. To produce full-length cDNA products for the viral genomic RNA, a set of LSV-specific primers could be designed based on the obtained sequence in this study and the known sequences of 3'-terminal region for the virus. Full-length cDNA copies of LSV, an 8.4 kb long, were directly amplified by the long-template RT-PCR technique from the purified viral genomic RNA samples. This full-length cDNA copies were analyzed by restriction mapping. The molecules produced in this study can be useful for the production of in vitro infectious cDNA clone, as well as, for the completion of genomic RNA sequence and genome structure for the virus.

• 한국식품영양과학회지
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• 제31권4호
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• pp.594-598
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• 2002

과일의 방사선 조사 여부 확인과 저장에 따른 변화를 측정하기 위해 감마선 조사로 유도된 DNA손상을 comet assay로 확인하였다. 키위, 오렌지, 배를 구입하여 0.1, 0.3, 0.5, 0.7, 1.0 kGy의 저 선량으로 조사하고 비 조사 시료와 조사 시료간의 DNA 손상정도를 tail length와 tail moment로 측정하였다. 과일 씨의 DNA를 형광 염색하여 이미지 분석기를 이용하여 comet 양상을 관찰한 결과, 모든 시료에서 비 조사 시료보다 조사시료의 tail length가 더 길었으며 조사 선량이 증가할수록 tail length가 유의적으로 길게 나타났다. Tail moment로 나타난 결과도 이와 비슷하였으나 전체적으로 tail length에 비해 그 민감도가 낮았다. 방사선 조사한 과일을 저온에서 3개월 동안 저장한 후에도 저장 전과 마찬가지로 모든 시료에서 비 조사시료보다 조사 시료의 tail length가 더 길었으며, 조사 선량이 증가할수록 tail length가 길게 나타나, 저장 후에도 comet assay를 이용해 조사 시료와 조사 시료의 방사선 조사여부를 검지할 수 있었다 따라서 본 연구결과 comet assay는 신선한 과일과 일정기간 저장한 과일의 방사선 조사여부 판별에 유용하게 사용될 수 있음을 알 수 있었다.

• KSII Transactions on Internet and Information Systems (TIIS)
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• 제15권8호
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• pp.2923-2943
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• 2021

The double helix structure of DNA makes it diverse, stable and can store information with high density, and these characteristics are consistent with the requirements of molecular communication for transport carriers. In this paper, a specific structure of molecular communication system based on DNA length coding is proposed. Transmitter (Tx) adopts the multi-layer golden foil design to control the release of DNA molecules of different lengths accurately, and receiver (Rx) adopts an effective and sensitive design of nanopore, and the biological information can be converted to the electric signal at Rx. The effect of some key factors, e.g., the length of time slot, transmission distance, the number of releasing molecules, the priori probability, on channel capacity is demonstrated exhaustively. Moreover, we also compare the transmission capacity of DNA-based molecular communication (DNA-MC) system and concentration-based molecular communication (MC) system under the same parameter setting, and the peak value of capacity of DNA-MC system can achieve 0.08 bps, while the capacity of MC system remains 0.025 bps. The simulation results show that DNA-MC system has obvious advantages over MC system in saving molecular resources and improving transmission stability.

• 대한의생명과학회지
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• 제7권1호
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• pp.1-5
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• 2001

A cDNA of coagulation Factor IX gene has been screened from the $\lambda$gt11 human fetal liver cDNA library, and used to construct a 2.8-kb full length cDNA after recombining with the N-terminal fragment from pTZ-FIX. Human genomic DNA was isolated, digested with the restriction endonucleases, TaqI, EcoRI, and HindIII, and Southern hybridization was performed using the full length factor IX cDNA as a probe. The hybridized bands generated by the restriction endonucleases were the followings: TaqI, 0.3, 1.0, 1.6, 1.8, 2.7, 3.7, and 5.3 kb bands; EcoRI, 1.8, 4.8, 4.9, 5.5, 6.8, and 12.6 kb bands; HindIII, 4.1, 4.4, 5.2, 5.8, 7.6, and 12.5 kb bands. When the Southern bands were physically mapped along the genome, about 50-kb continuous region harboring almost all of the genomic region of Factor Ⅸ gene was covered. These results suggest a possibility of using an exonal cDNA probe to diagnose abnormalities including large deletions, insertions, and rearrangements along the genome, if there is any.

• 한국식생활문화학회지
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• 제19권3호
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• pp.251-258
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• 2004

The changes in DNA damage were investigated during storage after irradiation. Potato, garlic were irradiated at 0.05, 0.07, 0.1 and 0.15 kGy and stored for 3 months. Ginger was irradiated at 0.01, 0.02, 0.03, 0.04 and 0.05 kGy and stored for 1 month. The comet assay was applied to the sample immediately after irradiation and at the end of storage. Samples were isolated, grounded and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were electrophoresed for 1 min. and then stained. DNA fragmentation in seeds caused by irradiation was quantified as tail length and tail moment (tail length ${\times}%$ DNA in tail) by comet image analyzing system. Right after irradiation, the differences in tail length between unirradiated and irradiated samples were significant(p<0.05) in potato, garlic and ginger. With increasing the irradiation doses, statistically significant longer extension of the DNA from the nucleus toward anode was observed. The results represented as tail moment showed similar tendency to those of tail length. Similarly in the stored samples, even 1 or 3 months after irradiation, all the irradiated samples significantly showed longer tail length than the unirradiated controls. These results indicate that the comet assay could be one of the simple methods of detecting irradiated samples. Moreover, the method could detect DNA damage even after 1 or 3 months after irradiation.

• 생명과학회지
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• 제23권10호
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• pp.1260-1266
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• 2013

본 연구에서는 H. marmoreus 3-10균주와 H. marmoreus 1-1균주의 ribosomal DNA (rDNA) cluster의 분석이 수행되었다. Small subunit (SSU)와 intergenic spacer 2 (IGS 2)는 부분적으로 염기서열이 결정되었고, internal transcribed spacer 1 (ITS 1), 5.8S, internal transcribed spacer 2 (ITS 2), large subunit (LSU), intergenic spacer 1 (IGS 1), 5S는 완전하게 염기서열이 결정 되었다. 팽이버섯 H. marmoreus 3-10균주와 H. marmoreus 1-1균주의 rDNA cluster는 총 7,049 bp로 결정되었다. SSU은 1,796 bp, ITS1은 229 bp, 5.8S은 153 bp, ITS2는 223 bp, LSU은 3,348 bp, IGS1은 390 bp, IGS2은 900 bp로 염기서열이 분석되었다. 결정된 rDNA cluster의 총 7,049 bp 중에서 17 bp가 다름이 확인되었고, 각각 SSU (2 bp), ITS (3 bp), LSU (9 bp), IGS (3 bp)에서 차이를 확인하였다. ITS regions의 2차 구조 결과 5개의 stem-loop가 있음이 드러났다. 흥미롭게도, 이들 stem-loop 사이에서 stem-loop V에서 한 개의 상이한 염기가 다른 2차 구조를 나타냄을 확인하였다.

• Bulletin of the Korean Chemical Society
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• 제24권7호
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• pp.943-947
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• 2003

We have studied the migration behavior of single-stranded DNA using capillary gel electrophoresis under various conditions. It was found that optimum electric fields should be less than 150 V/cm for the good tradeoff between the separation time and the resolution. It seems that the gel matrix with the combination of different polymer average molecular weights is important to extend the maximum readable DNA bases. The total gel concentration less than 3.1% in the mixed gel system showed good separation efficiency up to 600 bases. The best result was obtained with the poy(ethylene)oxide (PEO) gel concentration of 1.2% of Mr 8,000,000 and 1.8% of Mr 600,000. We observed that the capillary length between 50 cm to 100 cm (effective length) should be employed for the optimization between the total DNA migration time and the maximum readable length. A trizma base-boric acid-ethlyenediaminetetraacetic acid (EDTA) (TBE) buffer was commonly used for DNA sequencing, but we found that 3-[tris(hydroxymethyl)methyl amino]-1-propane sulfonic acid (TAPS) buffer worked as well for the single-stranded DNA separation. Especially, TAPS buffer showed a good resolution for very short DNA bases (1 to 30 bases).

• 한국식품과학회지
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• 제31권4호
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• pp.906-911
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• 1999

저선량으로 조사된 곡류의 방사선 조사 여부를 판별 하는데 DNA 'Comet Assay'의 활용이 가능한지를 검토하였다. 참깨, 들깨, 밀, 보리 및 쌀을 0, 0.1, 0.3, 0.5, 0.7, 1.0 kGy로 조사하여 시료로 사용하였다. 시료에 PBS 용액을 가하여 세포 현탁액을 조제하고 agarose gel과 혼합하여 microscope slide 상에서 lysis한 후 전기영동하여 염색한 다음 DNA 절편의 이동에 의해 형성된 ‘comet’의 tail length를 측정하여 분석하였다. 0.3 kGy 이상으로 조사한 모든 시료에서 tail length가 비조사한 시료와 차이를 나타내어 조사 여부를 확실히 판별할 수 있었다. 또한 비조사된 시료에서는 tail이 없거나 아주 짧은 tail을 가진 세포들이 관찰되었으며, 조사된 시료에서는 조사선량의 증가에 따라 DNA의 손상정도가 증가함으로서 tail length가 길어지는 것이 관찰되었다. 특히 참깨, 들깨 및 밀의 경우는 0.1 kGy의 낮은 조사선량에서도 현미경을 통하여 육안으로 비조사한 시료와의 구별이 가능하였다. 따라서 DNA 'Comet Assay'는 간단하고 신속하며 비교적 저렴하게 실시할 수 있는 곡류의 방사선 조사 여부를 판별하는 데 활용할 수 있음을 확인하였다.