• 대한수의학회지
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• 제47권4호
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• pp.457-460
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• 2007

The objective of the study was to establish routine parentage testing system in Beagle dogs using 22 ISAG (International Society for Animal Genetics) canine microsatellite markers (2005). Blood collections were obtained from a mother dog, 4 candidate father dogs and 3 offspring (n = 8). Genomic DNA samples were extracted from 8 Beagle dogs blood for PCR analysis. PCR products for the allele were analyzed by ABI 3130 DNA Sequencer and GeneScan (Ver 3.0) analysis and Genotyper (Ver. 2.1) software. The genetic relationship of mother and 3 offspring as well as one father dog among 4 candidate father dogs was confirmed by microsatellite allele analysis. The results of locus for amelogenin, which was designed for sexing, were matching with real gender among 8 Beagle dogs (female; 217/217 homozygosity, male; 179/217 heterozygosity). Twenty two ISAG microsatellite markers are useful the parentage test of Beagle dogs. In addition, amelogenin is an applicable marker to detecting real sex in dogs.

• Journal of Oral Medicine and Pain
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• 제20권1호
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• pp.229-246
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• 1995

The deoxyribonucleic acid(DNA) was isolated from the pulp, dentin and enamel of the 4 fresh teeth and the 7 teeth left in room temperature for 10 years. Then it was examed to find out the usefulness for forensic dental medicine. Samples of the tooth-derived DNA amplified by polymerase chain reaction(PCR), electrophosed for sex determination by detection of X-Y homologous amelogenin gene and D1S80 locus detection. The following results have been achieved. DNA extraction was possible in pulp and dentin of the fresh teeth, so it could be applicatable to detection of X-Y homologous amelogenin gene for sex determination, amplification of D1S80 locus by PCR. Sex determination was possible in pulp and dentin of the teeth left at room temperature for 10 years. Also, possible the detection fo AMP-FLPs to increase PCR cycling up to 40. DNA was isolated from all pulp of the fresh teeth and the teeth left in room temperature for 10 years, and also isolated from the dentin of the fresh teeth, partially isolated(3/7) from the dentin of the teeth left in room temperature for 10 years, but DNA was not isolated from enamel. From the above investigation, DNA extraction, sex determination, amplification of D1S80 locus were successfully accomplished even though the teeth were left for 10 years at room temperature. Therefore, teeth, especially pulp, are highly reliable and applicable as molecular biological samples for individual identification.

• Journal of Oral Medicine and Pain
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• 제19권2호
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• pp.205-219
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• 1994

Cigarette butts from 5 smokers were gathered and then, placed in room temperature for 1, 3, 5, 7, 15 days. The possible use of the cigarette butts for individual identification was evaluated in sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene from the extracted DNA. 1. DNA extraction was possible in cigarette butts weree left in room temperature for 15days, so it can be applicatable to individual identification by polymerase chain reaction(PCR). 2. Amplification of X-Y homologous amelogenin gene by PCR made it possible to identify the sex in saliva stains (cigarette butts). 3. Amplification of D1S80 locus can be acquired from adding the boving serum albumin and hot start PCR procedures from forensic samples such as saliva stains (cigarette butts), so the AMP-FLPs examining is possible. 4. Genotype could be determined simply and rapidly using Amplitype$TM$ HLA-DQ$\alpha$ forensic kit in examining the HLA-DQA1 gene. From the investigation, DNA extraction, sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene was successfully done even though the cigarette butts were left for 15 days at room temperature. Therefore cigarette butts are highly reliable and applicatable as molecular biologic samples for individual identification.

• 치위생과학회지
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• 제12권5호
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• pp.486-492
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• 2012

치아 발생 동안 법랑질모세포에서의 Dynamin II 단백질 발현 강도는 출생 후 1일째 생쥐에 비해 출생 후 3일째와 출생 후 5일째 생쥐에서 각각 48%와 50%로 유의성 있게 증가하였으나, 출생 후 7일째와 출생 후 10일째 생쥐에서는 출생후 1일째의 생쥐에 비해 각각 16%와 12%로 유의성 있게 감소하였다. 이 결과로부터 Dynamin II가 분비법랑질모세포에서 형성되는 법랑기질 단백질인 amelogenin, ameloblastin, enamelin과 MMP-20 등과의 분비과립 수송에 연관됨이 보여졌다. Dynamin II는 치아 발생과정 중 발현되는 다양한 법랑기질을 구성하는 단백질을 포함하는 분비소포 형성을 촉진함으로써 과립의 수송에 관여할 것이라 생각되며 법랑질모세포에서의 법랑기질 단백질의 분비조절 가능성이 있음이 보여졌다. 따라서 Dynamin II를 통한 치주질환을 위한 유전자 치료와 법랑질 혹은 상아질의 유실 부분의 재생산의 자극과 촉진에도 임상적으로 사용될 수 있는 가능성이 있음이 보여졌다.

• 보존과학연구
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• 통권29호
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• pp.137-148
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• 2008

고고 유적지에서 출토된 뼈에서 추출한 DNA는 부식산과 풀빅산 등의 토양성분과 콜라겐 등 다양한 오염물질이 포함되어 있어 DNA의 추출 및 분석이 매우 어렵다. 본 연구에서는 phenol 추출법, silica 추출법 등 두 가지 대표적인 고대 DNA 추출법의 효율을 DNA 증폭 결과에 의하여 비교하였다. 또한 울트라급의 초음파를 시료 용해에 적용한 후 silica 추출법으로 DNA를 분리한 방법이 기존의 phenol 및 silica 추출법에 비하여 미토콘드리아 DNA와 아밀로제닌 유전자 증폭 결과가 더 우수한 것으로 나타났다.

• Journal of Oral Medicine and Pain
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• 제20권2호
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• pp.497-513
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• 1995

Human genomic Deoxyribonucleic acid(DNA) was extracted from teeth by boiling, salting-out, phenol, boiling-phenol methods. The author compared DNA concentration and its purity, the accuracy of sex determination and the results of the D1S80 locus detection among above 4 methods. The following results were obtained : 1. DNA concentration was the highest in pulp with salting-out method and DNA purity was higher in pulp with salting-out and phenol methods than other 2 methods. 2. Sex determination was possible using of the pulp and the dentin of the teeth with four methods but, it was impossible in the enamel and some pulp with boiling method. 3. Amplification of D1S80 locus occurred from pulp and dentin with salting-out, phenol, and boiling-phenol methods. 4. There are no differences among the amplification of X-Y homologus amelogenin gene by application of 4 methods and salting-out, phenol methods efficiently makes available to amplification of D1S80 locus. From the investigation DNA extraction, sex determination, amplification of D1S80 locus was successfully accomplished with salting-out, phenol, boiling-phenol methods Therefore above 3 methods are available and applicable as forensic odontology for individual identification.

• 보존과학회지
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• 제26권1호
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• pp.95-107
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• 2010

본 연구는 충청남도 서천군 옥남리 일대(갓재골, 우아실) 조선시대 회곽묘에서 출토된 4개체의 인골을 대상으로 조직학, 분자유전학, 골화학 분석 등 종합적인 생화학적 연구를 수행한 결과이다. 실체현미경과 주사전자현미경 분석에 의한 대퇴골의 조직학적 보존 상태는 매우 양호 하였으며, 생화학적 분석이 가능한 것으로 평가되었다. 아밀로제닌 유전자 분석과 미토콘드리아 DNA에 대한 피장자들의 모계 계통형 분석 결과 여성 1명은 B4a에 속하는 것으로 밝혀졌으며, 다른 여성 1명과 남성 2명의 피장자는 하플로그룹 D4b1으로 나타났는데 이들 3명은 가까운 모계 혈연관계 가능성이 있는 것으로 추정되었다. 갓재골에 합장된 두 피장자는 전통적인 매장방식으로 볼 때 부부합장묘인 것으로 추정되었다. 콜라겐의 탄소-질소 안정동위원소 분석 결과 피장자들은 주식으로 쌀, 보리, 콩 등의 $C_3$ 식물을 섭취했던 것으로 나타났다. 본 연구를 통하여 조선시대 회곽묘를 조성할 수 있었던 경제적 계층의 유전학적 특징, 관습적인 매장방식, 식생활 정보를 규명하였다. 이러한 결과는 조선시대 출토 인골에 대한 생화학적 연구의 잠재적인 가치와 중요성에 대해 시사하고 있다.

• International Journal of Oral Biology
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• 제32권1호
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• pp.13-22
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• 2007

The stem cell research is emerging as a cutting edge topic for a new treatment for many chronic diseases. Recently, dental stem cell would be possible for regeneration of tooth itself as well as periodontal tissue. However, the study of the cell characterization is scarce. Therefore, we performed the genetic profiling and the characterization of mouse fetus/neonate derived dental tissue and cell to find the identification during dental development. We separated dental arch from mandibles of 14.5 d fetal mice and neonate 0 d under the stereoscope, and isolated dental cells primarily from the tissues. Then, we examined morphology and the gene expression profiles of the primary cells and dental tissues from fetus/neonate and adult with RT-PCR. Primary dental cells showed heterogeneous but the majority was shown as fibroblast-like morphology. The change of population doubling time levels (PDLs) showed that the primary dental cells have growth potential and could be expanded under our culture conditions without reduction of growth rate. Immunocytochemical and flow cytometric analyses were performed to characterize the primary dental cell populations from both of fetus (E14.5) and neonate. Alpha smooth muscle actin (${\alpha}-SMA$), vimentin, and von Willebrand factor showed strong expression, but desmin positive cells were not detected in the primary dental cells. Most of the markers were not uniformly expressed, but found in subsets of cells, indicating that the primary dental cell population is heterogeneous, and characteristics of the populations were changed during culture period. And mesenchymal stem cell markers were highly expressed. Gene expression profile showed Wnt family and its related signaling molecules, growth factors, transcription factors and tooth specific molecules were expressed both fetal and neonatal tissue. The tooth specific genes (enamelin, amelogenin, and DSPP) only expressed in neonate and adult stage. These expression patterns appeared same as primary fetal and neonatal cells. In this study we isolated primary cells from whole mandible of fetal and neonatal mice. And we investigated the characteristics of the primary cells and the profile of gene expressions, which are involved in epithelial-mesenchymal interactions during tooth development. Taken together, the primary dental cells in early passages or fetal and neonatal mandibles could be useful stem cell resources.

• Molecules and Cells
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• 제26권3호
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• pp.314-318
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• 2008

Korean long-tailed goral (Nemorhaedus caudatus) is one of the most endangered species in South Korea. However, detailed species distribution and sex ratio data on the elusive goral are still lacking due to difficulty of identification of the species and sex in the field. The primary aim of this study was to develop an economical PCR-RFLP method to identify species using invasive or non-invasive samples from five Korean ungulates: goral (N. caudatus), roe deer (Capreolus pygargus), feral goat (Capra hircus), water deer (Hydropotes inermis) and musk deer (Moschus moschiferus). The secondary aim was to find more efficient molecular sexing techniques that may be applied to invasive or non-invasive samples of ungulate species. We successfully utilized PCR-RFLP of partial mitochondrial cytochrome b gene (376 bp) for species identification, and sex-specific amplification of ZFX/Y and AMELX/Y genes for sexing. Three species (goral, goat and water deer) showed distinctive band patterns by using three restriction enzymes (Xbal, Stul or Sspl). Three different sexing primer sets (LGL331/335 for ZFX/Y gene; SE47/48 or SE47/53 for AMELX/Y gene) produced sex-specific band patterns in goral, goat and roe deer. Our results suggest that the molecular analyses of non-invasive samples might provide us with potential tools for the further genetic and ecological study of Korean goral and related species.

• Journal of Oral Medicine and Pain
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• 제21권2호
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• pp.225-242
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• 1996

The author studied to determine the sex and age of 8 ancient human skeletal remains, which had excavated from ancient tombs located in Yeanri, Kimhae. Some kinds of personal identification methods their skulls and teeth were used for this study and the results were obtained as follows : 1. Sex determination was possible in ancient teeth from 4th to 5th century, using detection of X-Y homologous amelogenin gene by polymerase chain reaction. 2. DNA analysis proved that the materials examined were all male, but which always did not coincide with the results from other methods for sex determination including comparison of sexual differentiation of cranium and teeth and use of discriminant functions in the dental measurement. 3. There was little difference of the estimated ages between the methods by regression of pulp cavity and attrition in teeth. The ages from these two methods always did not coincide with ones from evaluation closure of cranial and palatal sutures. 4. Sex and age of the materials were determined as follows ; $\cdot$ Y9 was estimated to be male in early sixties. $\cdot$ Yl2 was estimated to be male in late twenties to early thirties. $\cdot$ Y37 was estimated to be male in early forties. $\cdot$ Y70 was estimated to be male in early sixties. $\cdot$ Y87 was estimated to be male in late forties. $\cdot$ Y109 was estimated to be male in early forties. $\cdot$ Yl29 was estimated to be male in late thirties to early forties. $\cdot$ Yl42 was estimated to be male in late fifties to early sixties.