• Title/Summary/Keyword: Amelogenin

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Sex Ratio Determination by Quantitative Real Time PCR using Amelogenin Gene in Porcine Sperm

  • Hwang, You-Jin;Bae, Mun-Sook;Yang, Jae-Hun;Kim, Bo-Kyoung;Kim, Sang-Ok;Lee, Eun-Soo;Choi, Sun-Gyu;Kwon, Ye-Ri;Seo, Min-Hae;Park, Choon-Keun;Kim, Dae-Young
    • Journal of Embryo Transfer
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    • v.24 no.3
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    • pp.225-230
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    • 2009
  • Sex-sorting of sperm is an assisted reproductive technology (ART) used by the livestock industry for the mass production of animals of a desired sex. The standard method for sorting sperm is the detection of DNA content differences between X and Y chromosome-bearing sperm by flow cytometry. However, this method has variable efficiency and therefore requires verification by a second method. We have developed a sex determination method based on quantitative real-time polymerase chain reaction (qPCR) of the porcine amelogenin (AMEL) gene. The AMEL gene is present on both the X and the Y chromosome, but the length and sequence of its noncoding regions differ between the X and Y chromosomes. By measuring the threshold cycle (Ct) of qPCR, we were able to calculate the relative frequency of X chromosome. Two sets of AMEL primers were used in these studies. One set (AME) targeted AMEL gene sequences present in both X and Y chromosome, but produced PCR products of different lengths for each chromosome. The other set (AXR) bound to AMEL gene sequences present on the X chromosome but absent esholthe Y-chromosome. Relative product levels were calculated by normalizing the AXR fluorescence to the AME fluorescence. The AMEL method accurately predicted the sex ratios of boar sperm, demonstrating that it has potential value as a sex determination method.

AUTOTRANSPLANTATION OF AN IMPACTED MAXILLARY PREMOLAR USING ENAMEL MATRIX DERIVATIVE: A CASE REPORT (역위 매복된 상악 소구치의 Enamel matrix derivative를 이용한 자가이식 치험례)

  • Oh, You-Hyang;Lee, Nan-Young;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.3
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    • pp.471-476
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    • 2003
  • The success of autotransplantation depends on the viability of periodontal ligament in the transplanted tooth. Mechanical injury to periodontal tissues frequently results in dental root resorption and ankylosis, which leads to the failure of transplantation. Enamel matrix derivative(EMD) Which contains several enamel matrix protein (amelogenin family) has been reported to be effective in some periodontal therapies has been recently used to induce periodontal regeneration. EMD promotes proliferation of periodontal ligament cells and is suggested to be useful for transplantation. In this case, we report a clinical case of EMD application in the transplantation of an impacted and immature tooth of a 14 year-old girl to enhance the periodontal regeneration.

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Individual Identification by Analysis of DNA from the Teeth (치아에서의 DNA 분석에 의한 개인식별)

  • Chang-Lyuk Yoon;Chong-Youl Kim
    • Journal of Oral Medicine and Pain
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    • v.20 no.1
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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.

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Mitochondrial DNA Analysis of Human Skeletal Remains Excavated from Myungam-ri site in Asan, Korea (아산시 명암리 출토 인골의 미토콘드리아 DNA 분석)

  • Kim, Yun-Ji;Kim, Sue-Hoon;Cho, Eun-Min;Lee, Jeong-won
    • 보존과학연구
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    • s.36
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    • pp.33-48
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    • 2015
  • In this study, ancient DNA analyses were carried out on the human skeletal remains from a historical cemetery site in Myeongam-ri, Asan, Korea. Human remains of 27 individuals out of tombs from the Goryeo to Joseon Dynasty were selected for the analysis of this study. In order to identify the genealogy of the population and traditional burial pattern of the cemetery, we conducted comparative analyses of the hyper variable regions (HVRs) in mitochondrial DNA (mtDNA) of each sample. We sequenced 9 segmental amplicons of HVRs and assigned relevant haplogroups according to the sequence polymorphism on the basis of the known mtDNA database. As a result, we were analyses 18 human remains of 27 individuals and result of amelogenin analysis were only 4 samples.

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Expression of Dynamin II in Ameloblast during Mouse Tooth Development (생쥐 치아 발생과정 중 법랑질모세포에서 Dynamin II 발현)

  • Choi, Jung-Mi;Moon, Deog-Hwan;Lee, Jung-Hwa
    • Journal of dental hygiene science
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    • v.12 no.5
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    • pp.486-492
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    • 2012
  • Immunostaing intensity of Dynamin II protein in ameloblast during mouse tooth development showed a significant increase of 48% at the postnatal day 3 and a significant increase of 50% at the postnatal day 5 as compared with the postnatal day 1, but showed a significant decrease of 16% at the postnatal day 7 and a significant decrease of 12% at the postnatal day 10 as compared with the postnatal day 1. From the above results, Dynamin II had relevance to secretion of amelogenin, ameloblastin, enamelin and matrix metalloproteinase-20 proteins for enamel formation in ameloblast. Dynamin II may be involved in the transport of vesicles containing proteins for enamel formation through the acceleration of vesicular formation and may be had a good possibility of secretory regulation of proteins for enamel formation in ameloblast. Therefore, Dynamin II have potential for being used in the field of gene theraphy for periodontal disease and in the regeneration for enamel and dentin tissues lost to dental caries.

Genetic Characteristics of mtDNA and STR marker in Human Bone Excavated from Mokgam-dong, Siheung in Korea (시흥 목감동 출토 인골의 미토콘드리아 DNA와 STR의 유전적 특징)

  • Seo, Min-Seok;Chung, Yong-Jae;Lee, Kyu-Shik;Park, Ki-Won
    • 보존과학연구
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    • s.24
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    • pp.153-167
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    • 2003
  • We performed nuclear DNA typing and mitochondrial DNA sequencing analysis based on PCR from an ancient Korean remainsexcavated from Siheung in Korea. 7 bones were collected and partially STR(short tandem repeat) systems, Sex determination Amelogenin kit(Promega co, USA), were used in this study. Mitochondrial DNAs were also amplified and sequenced by ABI 310 DNA sequencer. We know that sample no. 2 and no. 3 were females and also sample no. 2 and no.7 possessed the same maternal inheritance by mitochondrial DNA sequencing results. Throughout this research, the mitochondrial DNA sequencing of human in the middle of Joseon Dynasty in Korea is obtained. In addition, this finding will be an important foundation for the future research.

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Molecular Genetic and Histological Analysis for the Excavated Human Bone (출토 인골의 분자유전학 및 조직학적 분석연구)

  • Jee, Sang-hyun;Park, Ji-hee;Seo, Min-seok;Hong, Jong-ouk;Chung, Yong-jae
    • 보존과학연구
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    • s.28
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    • pp.75-90
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    • 2007
  • The ancient biomolecular remains are the potential source for paleobiology and paleoanthropology. Especially, ancient human specimens such as bone, teeth, and hair are powerful materials to identify historical origin and migration of ancestor population from the past. However, most excavated human specimens in archaeological sites have commonly problems as natural damage and exogenous contamination. We carried out histological and molecular analyses of excavated bone from the historic sites in South Korea from the recently discovered in tumulus of Seochun and Naju. Biological deterioration of bone was observed anatomically by optical and scanning electron microscope (SEM). We extracted degraded DNA, and amplified hyper variable region (HVR) of mitochondrial DNA (mtDNA) and amelogenin of nucleus DNA. This study applied and examined the relationships between histological preservation and DNA survival in excavated bone.

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Usefulness of Biochemical Analysis for Human Skeletal Remains Assigned to the Joseon Dynasty in Oknam-ri Site in Seocheon, Korea (조선시대 인골에 대한 생화학적 분석의 유용성: 서천군 옥남리 회곽묘 출토 인골을 중심으로)

  • Kang, So-Yeong;Kwon, Eun-Sil;Moon, Eun-Jung;Cho, Eun-Min;Seo, Min-Seok;Kim, Yun-Ji;Jee, Sang-Hyun
    • Journal of Conservation Science
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    • v.26 no.1
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    • pp.95-107
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    • 2010
  • Biochemical research was carried out on 4 human skeletal remains from historical lime-layered tombs assigned to the Joseon Dynasty in Oknam-ri, Seocheon. The preservation of femur was evaluated by stereoscopic microscopy and scanning electron microscopy. Most of specimens showed good histological preservation. The histological results proved to be a good potentiality for biochemical analysis using bio-molecules. The amelogenin gene and mitochondrial DNA (mtDNA) analyses revealed that three specimens perhaps have maternal consanguinity due to sharing with mtDNA haplogroup D4b1, and two specimens buried in the same tomb were a couple in Gatjaegol site. Carbon and nitrogen stable isotope analysis indicated that four deads diet were built around C3 plant as rice, barley, wheat and bean. In this study we characterized genetic and diet features from the social stratum who could make lime-layered tombs during period of the Joseon Dynasty. The results suggest that biochemical research using the human skeletal remains from the Joseon Dynasty has the great potential and reasonable value for archaeology, anthropology, and population genetics.

Personal identification of the excavated ancient human bone through molecular-biological methods (분자생물학적 방법을 통한 출토인골의 개인 동정-사천 늑도 출토 인골과 민통선 민묘 출토 인골을 중심으로)

  • Seo, Min-Seok;Lee, Kyu-Shik;Chung, Yong-Jae;Lee, Myeong-Hui
    • 보존과학연구
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    • s.22
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    • pp.27-40
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    • 2001
  • DNA typing is often used to determine identity from human remains. Recently, the molecular biological analysis of ancient deposits has become possible since methods for the recovery of DNA conserved in bones or teeth from archaeological remains have been developed. In the field of archaeology, one of the most promising approaches is to identify the individuals present in a mass burial site. We performed nuclear DNA typing and mitochondrial DNA sequencing analysis based on PCR from a Korea ancient human remain excavated from Sa-chon Nuk-island and civilian access controlline(CACL). A femur bone were collected and successfully subjected to DNA extraction, quantification, PCR amplification, and subsequently typed for several shot tandem repeat(STR)loci. 4 types of STR systems used in this study were CTT multiplex(CSF1PO, TPOX, TH01), FFv multiplex(F13A01, FESFPS, vWA), Silver STRⅢ multiplex(D16S539, D7S820, D13S317), and amelogenin for sex determination. This studies are primarily concerned with the extraction, amplification, and DNA typing of ancient human bone DNA samples. Also, it is suggestive of importance about closely relationship between both fields of archaeology and molecular biology.

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