Diamond™ Nucleic Acid Dye를 사용한 Touch DNA 시각화 및 검출

오대환(Oh, Dae Hwan),홍승범(Hong, Seung Bum),최하영(Choi, Ha Young),신강일(Shin, Kang Il) 경찰대학 경찰학연구편집위원회 2021 경찰학연구 Vol.21 No.3

본 연구에서는 범죄현장 및 증거물에 잠복되어 있는 DNA (Deoxyribose Nucleic Acid)를 시각화하여 정확한 위치에서 유전자 증거물을 채취하고자 젤 전기영동에 사용되는 핵산 염색 시약 중 하나인 Diamond™ Nucleic Acid Dye를 사용하였다. 사람이 손으로 만진 물품의 표면에 Diamond™ Nucleic Acid Dye(10,000X) 원액을 75% 에탄올로 20배 희석하여 염색하였다. 생물학적으로 근연관계가 없는 4명의 공여자에게 멸균 처리된 물품을 만지게 한 후, 20배 희석한 DD(Diamond™ Nucleic Acid Dye)를 물품의 표면에 피펫으로 점적하여 Touch DNA를 염색하였다. 그리고 Dino-Lite 디지털형광현미경을 사용하여 물품 표면의 형광을 확인하였다. DD 분자는 구조적으로 DNA의 골격(sugar-phosphate backbone)에 효과적으로 결합할 수 있기 때문에 ssDNA(single-stranded DNA), dsDNA(double-stranded DNA) 그리고 RNA를 형광으로 확인 할 수 있다. DD는 dsDNA를 기준으로 청색 파장(494nm)에서 최대 여기(excitation)를 가지며 DNA와 결합하면 녹색 형광(558nm)을 방출(emission)한다. 이로써 4명의 공여자가 만진 물품에서 녹색 형광을 방출하는 발광체를 관찰할 수 있었으며, 멸균 면봉에 멸균수를 묻혀서 녹색 형광을 발현하는 발광체를 채취하였다. 현재 경찰청에서 국립과학수사연구원으로 유전자 감정 의뢰과정과 동일하게 사용 중인 유전자 증거물 채취 키트에 포장하여 한국유전자정보연구원으로 가져간 후 STR(Short Tandem Repeat) profiling 분석을 하였으며, 공여자 4명의 구강상피세포 DNA에서 분석된 STR profile을 대조한 결과 개인식별이 가능한 것을 확인하였다. 이 방법이 보급 된다면 보이지 않는 DNA를 시각화하여 유전자증거물을 채취할 수 있도록 과학수사관을 안내하고 DNA가 없는 시료의 무분별한 감정 처리 시간 단축 및 예산 절약의 효과가 있다고 사료되었다. This study used Diamond™ Nucleic Acid Dye, a nucleic acid staining solution used for gel electrophoresis to precisely locate and collect genetic evidence from the crime scene and evidence by visualizing DNA (Deoxyribose Nucleic Acid). Diamond™ Nucleic Acid Dye (10,000X) was diluted by 20times with 75% ethanol and was used to dye the surface of the touched items. After having four biologically unrelated donors touch sterilized items, the diluted Diamond™ Nucleic Acid Dye(DD) solution was dripped onto the surface of the items using a pipette to stain the Touch DNA. A dino-Lite digital fluorescence microscope was then used to observe fluorescence. ssDNA(single-stranded DNA), dsDNA(double-stranded DNA) and RNA can be confirmed with fluorescence since the structure of DD molecules effectively combine DD molecules with the sugar-phosphate backbone of the DNA. With dsDNA at the baseline, DD has the greatest excitation at the blue wavelength(494nm) and emits green fluorescence(558nm) when combined with DNA. Green fluorescence was observed on the items touched by the four donors, and sterilized cotton swabs moistened with sterile water were used to collect luminous matter emitting green fluorescence. To replicate the current genetic analysis request procedure, the sample was placed in the genetic evidence collection kit, sent to the Korea Genome Information Institute from the police agency, and analyzed with STR(Short Tandem Repeat) profiling. A comparison with the STR profiles derived from the buccal epithelial cells of the four donors confirmed that the collected sample could successfully identify the donors. Widespread use of the technique presented in this research is expected to visualize invisible DNA and guide forensic field agents, reduce the processing time of DNA-less staining solution and result in a budget-saving.

DNA 컴퓨팅과 진화 모델을 이용하여 Traveling Salesman Problem를 해결하기 위한 DNA 서열 생성 알고리즘

김은경(Eun-Gyeong Kim),이상용(Sang-Yang Lee) 한국지능시스템학회 2006 한국지능시스템학회논문지 Vol.16 No.2

현재 막대한 병렬성을 갖는 DNA 컴퓨팅을 이용하여 Traveling Salesman Problem (TSP)를 해결하기 위한 연구가 진행되고 있다. 하지만 기존의 방법은 그래프 문제의 표현에서 DNA의 특성을 고려하지 않아, 실제 생물학적 실험 결과와의 차이가 발생하고 있다. 따라서 DNA의 특성을 반영하고 생물학적 실험 오류를 줄일 수 있는 DNA 서열 생성 알고리즘이 필요하다. 본 논문에서는 DNA 컴퓨팅에 진화 모델의 하나인 DNA 코딩 방법을 적용한 DNA 서열 생성 알고리즘을 제안한다. 제안한 알고리즘은 TSP에 적용하여 기존에 단순 유전자 알고리즘과 비교하였다. 그 결과 제안한 알고리즘은 오류를 최소화한 우수한 서열을 생성하고 생물학적 실험 오류율도 줄일 수 있었다. Recently the research for Traveling Salesman Problem (TSP) using DNA computing with massive parallelism has been. However, there were difficulties in real biological experiments because the conventional method didn't reflect the precise characteristics of DNA when it express graph. Therefore, we need DNA sequence generation algorithm which can reflect DNA features and reduce biological experiment error. In this paper we proposed a DNA sequence generation algorithm that applied DNA coding method of evolution model to DNA computing. The algorithm was applied to TSP, and compared with a simple genetic algorithm. As a result, the algorithm could generate good sequences which minimize error and reduce the biologic experiment error rate.

DNA 칩 데이터의 분석을 위한 개미 집단 시스템 알고리즘 기반의 분류화 기법

이민수,이윤경,최옥주,맹보연 한국정보과학회 2008 데이타베이스 연구 Vol.24 No.3

DNA chips are widely used in gene analysis due to the fact that it can provide various information related to genes and experiments. The DNA chip is composed of a 2-dimensional array with a gene axis and a sample axis, and is much more efficient and practical than time consuming and costly biological experiments. A DNA chip analysis system extracts various data from DNA chips and inspects the data. Among the various methods to inspect the data, classification is one of the most popular methods. The classification method depends on the data types and characteristics. Considering that the DNA chip data is large, using an algorithm that imitates the biological system such as the ant algorithm is a highly desired approach. In this research the Ant Colony System Algorithm is used to discover classification rules from DNA chip data. DNA 칩은 최근 유전자 분석에서 유전자와 실험 등에 대한 다양한 정보를 쉽게 제공할 수 있어서 널리 사용되고 있다. 유전자(gene)와 실험(sample)의 두 축으로 이루어진 2차원의 DNA 칩은 결과를 얻기 위한 많은 비용과 시간을 필요로 하는 생물학적 실험보다 더 효율적이고 실용적인 장접 때문에 오늘날 널리 사용되고 있다. DNA 칩 분석 시스템은 다양한 종류의 DNA 칩에서 자료를 추출하고 유용한 정보를 얻기 위해 데이터를 조사한다. 데이터를 조사하는 방법은 다양하지만 일반적으로 사용되는 방법 중 하나가 분류화(classification)이다. 데이터를 분류하기 위한 알고리즘은 자료 유형이나 수많은 특성에 종속적인 것으로 바이오 칩 데이터가 매우 크다는 것을 고려하면 개미 알고리즘인 개미 집단 알고리즘(Ant Colony System Algorithm)을 사용ㅎ여 DNA 칩 데이터로부터 분류 규칙을 발견하는 기법을 제안하고 실험으로 검증하였다.

디엔에이(DNA)신원확인정보의 수집 및 관리에 관한 제문제

서주연 경찰대학 치안정책연구소 2015 치안정책연구 Vol.29 No.3

현대 과학기술의 발달은 범죄를 수사함에 있어 긍정적인 역할을 하기도 한다. 유죄입증을 위한 증거수집 및 분석에 있어서도 과거에는 관련기술의 부재로 증거로서의 중요성을 가지지 못했던 것들이 이제는 유죄의 입증에 중요한 역할을 하고 있다. 과거 혈액형의 동일성을 증명하였던 것에 불과하던 혈흔에서도 이제는 DNA유전자형 분석을 통해 범행을 입증할 수 있기에 이른 것이다. 이에 우리는 DNA정보가 가지는 중요성에 주안점을 두고 DNA정보 운용을 위한 디엔에이(DNA)신원확인 정보의 이용 및 보호에 관한 법률을 마련하여 시행중에 있다. 그러나 이와 관련해서는 DNA신원확인정보의 수집․관리 적용대상이 너무 포괄적이라는 지적에서부터 DNA대량검색을 통한 그물망 수사 문제 및 관련 정보의 폐기와 관련한 문제에 이르기 까지 여러 비판이 제기되고 있다. DNA정보 운용과 관련해서는 우리뿐만 아니라 세계 각국에서 관심을 가지고 있는 분야이며 그 중 미국의 경우 1990년대부터 관련 법제가 마련된 바 있다. 미국은 DNA관련 규정을 연방과 각 주에서 따로 규정하고 있으나 각 시스템이 연계되어 운용되고 있다는 특징이 있다. 뿐만 아니라 DNA신원확인정보의 대상을 확대하고 있으며 관련 정보의 폐기와 관련해서도 각 주별 관대한 입장을 취하고 있는 경우가 많아 우리와 차이를 보이고 있다. 이에 본고에서는 현재 실행 중인 디엔에이신원확인 정보의 이용 및 보호에 관한 법률에 대해 전반적으로 검토한 후 미국에서 시행 중인 DNA관련 법제와의 비교를 통해 우리 법제의 나아갈 방향에 대하여 고찰하였다. Development of science technology has a positive influence in crime investigation. Especially technology of DNA analysis gave us possibility to compare prisoners blood or body fluids. DNA information for identification is based on Deoxyribonucleic acid, it gave us to information for human identity. DNA analysis is a kind of forensic science, and it means a lot in police investigation today. DNA information raised very important thing to solve the case, so the DNA Act in Korea was enacted in 2010. It regulated method of collecting samples, warrant issues, disposal of it and delete the data etc. Nonetheless it has some problems like a including none violent crimes, collecting samples in suspects and massive collecting issues. United States is the one of the country has attention about DNA information and well regulated. Database about DNA information in US separated by CODIS and States databases, but its connected both way. It also has a feature like a expansion to none violent crime, regulations of saved DNA samples for the re-analysis, these things are huge differences between us. DNA information is very important in human life, so we have to have more caution to collecting and make databases.

DNA-dependent Protein Kinase Complex : a Multifunctional Protein in DNA Repair and Damage Checkpoint

Lee, Suk-Hee,Kim, Chung-Hui Korean Society of Molecular Biology 2002 Molecules and cells Vol.13 No.2

Potentiation of chemosensitivity in multidrug-resistant human leukemia CEM cells by inhibition of DNA-dependent protein kinase using wortmannin

Kim, Sun-Hee,Um, Jee-Hyun,Kim, Dong-Wan,Kwon, Byung-Hyun,Kim, Dong-Wan,Chung, Byung-Seon,Kang, Chi-Dug Institute of Genetic Engineering Changwon National 2000 Gene and Protein Vol.4 No.-

DNA-dependent protein kinase (DNA-PK) is activated by DNA stand breaks and participates in DNA repair. Its regulatory subunit, Ku autoantigen, binds to DNA and recruits the catalytic subunit (DNA-PKcs). We show here a new role of DNA-PK in the development of multidrug resistance (MDR). The Ku-DNA binding activity, the levels of Ku70/Ku80 and DNA-PKcs in MDR variants, CEM/VLB_(10-2). CEM/VLB_(55-8) and CEM/VLB_100 were higher than those in their parental drug-sensitive CEM cells in a drug resistance-dependent fashion. Also, CEM/VLB_100 cells showed about 3-fold increase of DNA-PK enzyme activity as compared with CEM cells. Similar results were observed in another MDR cell line, FM3A/M mouse mammary carcinoma cells. Moreover, we observed the CEM/VLB_100 cells were about 11-fold sensitive to wortmannin, which inhibits DNA-PK, compared with the CEM cells, and sensitized the MDR cells when combined with either bleomycin or vincristine, but have a little effect on CEM cells. Wortmannin was shown to inhibit DNA-PK and Ku-DNA binding activity of CEM/VLB_100 cells dose dependently but had a little or no effect on their parental cells. Our results suggested that enhanced expression of DNA-PK participates in the development of MDR, and the use of DNA-PK inhibitors such as wortmannin is likely to improve the effectiveness of anticancer drugs and thus could partially overcome drug resistance in DMR cells, through its ability to inhibit Ku/DNA-PK activity. ⓒ 2000 Elsevier Science Ltd. All rights reserved.

유방암에서 DNA 성상과 예후 및 임상병리적 상관관계에 대한 연구

감봉수,양영일,김상효 인제대학교 1994 仁濟醫學 Vol.15 No.3

본 연구의 목적은 Flow Cytometry(FCM)를 이용하여 유방암의 DNA양과 세포증식 도를 반영하는 DNA ploidy, 5-phase fraction을 측정하여 병기 및 전이와의 관계와 수술 후 재발의 위험도를 알 수 있는 예후추정 인자로서의 관련성을 알아보고자 하는데 있다. The nuclear DNA content has recently emerged as a new prognostic indicator in various cancers. Flow cytometry(FCM) has become a generally accepted analytical method, thousands of nuclei can be evaluated for their DNA content with a good resolution. Breast cancer in one of those malignant tumors which have been extensively investigated by flow cytometry. in recent, there has been reported that DNA ploidy is an important prognostic significance of breast cancer. The nuclear DNA contents of 32 female breast cancers were deter mined by flow cytometric technique using paraffin-embedded tissue. All patients were fort lowed-up far the period ranging from 3 months to 8 years and recurrence and survival period were identified. The mean Value of comfficient of variance in DNA histogram Was 6.8±1.0(3.8-8.0). DNA diploid tumor was twenty cases(62.5%) and nondiploid was twelve cases(37.5%). In patients wish diploid tumors, the mean S-phase fraction(SFP) was 17.3±7.9% in the bnondiploid tumors, the mean S-phase fraction(SPF) was 22.0±8.1%. DNA ploidy data was correlate with various clincal and histopathologic factors : the significant correlation existed between the tumor size, clinical stage, axillary lymph node or distant metastasis and DNA ploidy status(P <0.05). Based on these results, it is concluded that nuclear DNA ploidy status and SPF are an independent prognostic factor predicting for recurrence of the disease, and nondiploid breast cancer with high SPF needs more active postoperative adjuvant treatment because of its biologic aggressiveness. But additional longterm studies are necessary before tumor ploidy and SPF cna be used to select patient who should or shoud not receive systemic therapy.

Activation Mechanism of Protein Kinase B by DNA-dependent Protein Kinase Involved in the DNA Repair System

Li, Yuwen,Piao, Longzhen,Yang, Keum-Jin,Shin, Sang-Hee,Shin, Eul-Soon,Park, Kyung-Ah,Byun, Hee-Sun,Won, Min-Ho,Choi, Byung-Lyul,Lee, Hyun-Ji,Kim, Young-Rae,Hong, Jang-Hee,Hur, Gang-Min,Kim, Jeong-Lan Korean Society of ToxicologyKorea Environmental Mu 2008 Toxicological Research Vol.25 No.4

DNA-dependent protein kinase(DNA-PK) is involved in joining DNA double-strand breaks induced by ionizing radiation or V(D)J recombination and is activated by DNA ends and composed of a DNA binding subunit, Ku, and a catalytic subunit, DNA-PKcs. It has been suggested that DNA-PK might be $2^{nd}$ upstream kinase for protein kinase B(PKB). In this report, we showed that Ser473 phosphorylation in the hydrophobic-motif of PKB is blocked in DNA-PK knockout mouse embryonic fibroblast cells(MEFs) following insulin stimulation, while there is no effect on Ser473 phosphorylation in DNA-PK wild type MEF cells. The observation is further confirmed in human glioblastoma cells expressing a mutant form of DNA-PK(M059J) and a wild-type of DNA-PK(M059K), indicating that DNA-PK is indeed important for PKB activation. Furthermore, the treatment of cells with doxorubicin, DNA-damage inducing agent, leads to PKB phosphorylation on Ser473 in control MEF cells while there is no response in DNA-PK knockout MEF cells. Together, these results proposed that DNA-PK has a potential role in insulin signaling as well as DNA-repair signaling pathway.

Effect of Cobaltous Chloride on the Repair of UV-induced DNA Damage

Kim,Kug-Chan,Kim,Yung-Jin,Lee,Kang-Suk 대한방사선 방어학회 1995 방사선방어학회지 Vol.20 No.2

본 연구에서는 유전자 손상회복에 관여하는 단백질을 이용하여 돌연변이 생성을 억제시키는 물질로서 알려진 cobaltous chloride 가 유전자 손상회복에 미치는 영향을 연구하므로서 방사선으로 인한 손상방지 및 방사선 방어효과에 대한 적용가능성을 평가하였다. Cobaltous chloride가 RecA 단백질의 기능에 미치는 영향을 조사한 결과 RecA 단백질에 의한 DNA strand exchange 반응에 있어 cobaltous chloride처리로 RecA 단백질이 ssDNA로 부터 SSB 단백질과 더 효과적으로 경쟁함으로써 안정된 RecA-ssDNA complex 의 형성을 유도하고, 증가된 ATPase 활성에 의한 ATP 가수분해로 손상된 DNA의 회복이 촉진될 수 있다는 사실을 입증 해주고 있다. 또한 RecA 단백질은 UV에 의한 손상된 supercoiled DNA에 더 효과적으로 결합 됨이 관찰되었으며 UV 션량과도 상관관계가 있음을 확인하였다. 따라서 이와같은 연구겨러과들은 방사선으로 인한 유전적인 손상방지 및 방사선 방어효과에 관한 연구에 적용될 수 있을 것으로 기대된다. To develop methods to reduce radiation risk and apply such knowledge to improvement of radiation protection, the efrfects of cobaltous chloride known as bioantimutagen on the function of E. coli RecA protein involved in the repair of DNA damage were examined. The results demonstrated two distinct effects of cobaltous chloride on the RecA protein function necessary for the strand exchange reaction. Cobaltous chloride enhanced the ability of RecA protein to displace SSB protein from single-stranded DNA and the deplex DNA-dependent ATPase activity. RecA protein was preferentially bound with UV-irradiated supercoiled DNA as compared with nonirradiated DNA. The binding of RecA protein to UV-irradiated supercoiled DNA was enhanced in a does-depkendent manner. It is likely that studies on the factors affecting repair efficiency and the DNA repair proteins may provide information on the repair of ionizing radiation-induced DNA damage and the mechanism for DNA radioprotection.

DNA 채취제도와 채취영장의 법적성질과 그 쟁점

조광훈 법제처 2021 법제 Vol.692 No.-

DNA sampling system is for effective countermeasures against crimes that have become more and more ferocious and intellectual. The system is based on the Act on Use and Protection of DNA identification Information (hereinafter ‘DNA Act’). It is considered reasonable to regard the Clause 1, Article 5 of the DNA Act that regulates crimes subject to sampling as a non-mandatory provision. DNA sampling system shall be considered as a special disposition that administrative measure, compulsory measure and security measure are mixed. It is also considered as a system that the Risk Criminal Procedure Act is realized concretely as the system is placing emphasis on the operation to compare․analyze DNA preemptively and actively for preventing crimes. Although the system puts focus on prevention of crimes in both function of identification and the future regarding unsolved crimes that occurred in the past, it is necessary to look back whether or not the system limits basic human rights of subjects of sampling unnecessarily and to respect human and basic rights of subjects of sampling while emphasizing effectiveness of DNA sampling system. Sampling from the body of a prisoner or arrested suspect with DNA sampling warrant is rather close to the in personal compulsory measure, and sampling of specimen from the place of crime is close to the legal nature of in rem compulsory measure. Due to the characteristic, legal nature of DNA sampling warrant differs from verification warrant, detention center for judgment and permit for judgment. Collection of DNA shall be considered that it includes the process of identifying DNA. Therefore, an assertion that additional warrant for identification other than sampling warrant is not appropriate. And the assertion that a judge evaluates the risk and prediction of repeated crime while examining issuance of a sampling warrant is also inappropriate in a sense that the risk and prediction of repeated crime have already been evaluated as the Item 1 through 11 of Clause 1, Article 5 of the DNA Act regulates crimes subject to sampling. And as for sampling warrant, reasonableness․proportionality and crimes subject to sampling that are required for arrest warrant or search․seizure․verification warrant have already been stipulated on the DNA Act, so it is considered necessary to assume that such requirements are not included in the requirements for issuance of DNA sampling warrant. Accordingly, whether or not the subject of sampling accepted request of a sampling institute for DNA collection must be one and only requirement when a judge examines issuance of a sampling warrant.