• 제목/요약/키워드: Restriction pattern

검색결과 226건 처리시간 0.021초

체세포복제 소 배반포의 Oct-4 발현과 DNA 메틸화 변화 (Expression and DNA Methylation Change of Oct-4 in Cloned Bovine Blastocysts)

  • 차병현;최정상;황성수;정학재;임기순;양병철;김명직;조재현;성환후;고응규
    • 한국수정란이식학회지
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    • 제23권3호
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    • pp.133-139
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    • 2008
  • DNA methylation is one of the reasons for poor survival of clone animals. The OCT-4 gene is essential for maintaining pluripotency of embryonic stem (ES) cells and early embryos. We previously reported that the 5'-promoter region of Oct-4 gene was a target of DNA methylation and the methylation status was changed variously during embryonic development in bovine. The study conducted to examine the expression and methylation pattern of tissue-dependent differentially methylated region (T-DMR) of Oct-4 gene in bovine somatic cell nuclear transfer (SCNT) and in vitro fertilization (IVF) blastocysts. The Oct-4 gene expression was evaluated by RT-PCR and fluorescence immunocytochemistry. The methylation pattern of T-DMR was analyzed using restriction mapping and bisulfite sequencing methods. The Oct-4 transcripts were highly expressed in IVF, while they were not expressed in SCNT. The Oct-4 protein was not detected or expressed at very low level in SCNT, the intensity of Oct-4 protein, however, was strong in IVF. On the other hand, the T-DMR of Oct-4 gene was hypermethylated in SCNT compared to that of IVF. These results suggested that expression and the failure of demethylation of Oct-4 gene was closely associated with incomplete development of SCNT embryos.

Archicitrus와 Metacitrus로부터 Mitochondrial DNA의 분리 및 특성 (Isolation and Characterization of Mitochondrial DNA from Arehicityars and Metacitrus)

  • 이숙영;박민희
    • 한국식품영양학회지
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    • 제8권4호
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    • pp.307-317
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    • 1995
  • The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

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Expression and Characterization of Bovine DNA Methyltransferase I

  • Chang, Yoo-Min;Yang, Byoung-Chul;Hwang, Seong-Soo;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • 제33권2호
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    • pp.93-98
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    • 2009
  • In this study, bovine Dnmt1 cDNA was sequenced and detected Dnmt1 mRNA level in bovine tissues by northern blot, methylation pattern of genome by southern blot, specific localization of Dnmt1 in mouse and bovine preimplantation embryos by immunocytostaining and Dnmt1 protein level in ovary and testis by western blot. Bovine Dnmt1 cDNA sequence showed more homology with that of human than mouse and rat. The RNA level of Dnmt1 was 10 times higher expression in placenta than other tissues. This indicates that placenta was hypermethylated compared to others organs. The genomic DNA could not be cut by a specific restriction enzyme (HpaII) in placenta, lung and liver of bovine. It suggests that Dnmt1 in some somatic cells was already methylated. Dnmt1, which has the antibody epitope 1316~1616, was distributed in nucleus and cytoplasm including the stage of pronuclear stage and maturation of oocyte and gradually weaken to blastocyst stage compare to negative. In addition, Dnmt1 was strongly expressed in tetraploid embryo and cloned 8-cell than IVF 8-cell. An aberrant pattern of DNA methylation in cloned embryo may be abnormal development of fetus, embryonic lethality and placenta dysfunction. The somatic specific band (190kDa) was appeared in ovary and testis, but oocyte specific band (175kDa) was not. Further investigations are necessary to understand the complex links between the methyltransferases and the transcriptional activity of genes in the cloned bovine tissues.

일부 농촌지역 주민의 보건지소 이용 양상과 관련요인 (Rural Health subcenter Utilization Pattern and Its Related Factors)

  • 손석준
    • 농촌의학ㆍ지역보건
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    • 제19권2호
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    • pp.97-106
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    • 1994
  • In order to estimate the utilization pattern of a rural health subcenter, and to identify the recognition for it among the inhabitants in Kogsung district, a questionnaire survey was carried out for objects of 708 population. The results observed were as follows; 1. The annual utilization rate of a rural health subcenter for a basic health service unit was 27.5 per 100 persons, and annual mean visiting times was 1.43 times. 2. The most frequent disease by, annual health subcenter utilization illness was respiratory disease(26.5%), and the next was musculoskeletal disease(23.9%), gastrointestinal disease(15.9%) by order. 3. Favorite reasons for community health subcenter utilization were lower medical cost(23.4%), near distance from living place(20.7%) and lower disease severity(19.5%) by order. But disfavorite reasons for it were non effective treatment(26.2%), insufficient equipment(25.4%) and absence of specialist(17.4%) by order. 4. Insufficient items about community health subcenter utilization were restriction of treatment limit(47.1%), lower reliance(22.4%) and not punctral(21.8%) by order. 5. The results of logistic regression analysis suggested that statistically significant factors in health subcenter utilization were age, educational level and the nearest medical facility class. 6. There was no difference between recognition for a community health subcenter's work and actual utilizing service, and desirable works for it were disease preventing service, disease control of elderly and sanitation control by order. These results suggested that to increase the utilization of rural health subcenter and to promote the accessibility of rural residents to primary health care, there must be considered public relation about health subcenter, expansion of health equipment and recognition about access time.

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위 이형성 상피 병변의 클론성에 대한 분자병리학적 연구 (Clonality Assay of Dysplastic Epithelial Lesions of the Stomach)

  • 최호수;김미숙;박재우;박창수;김영진;정상우
    • Journal of Gastric Cancer
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    • 제1권3호
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    • pp.129-135
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    • 2001
  • Purpose: Dysplasia or flat adenoma of the stomach is regarded as a precancerous lesion. However, the frequency and the evolutionary process of malignant transformation of gastric dysplasia are still debated. In order to see whether the lesion was a monoclonal or a polyclonal proliferation, clonality was assayed by X-linked HUMARA polymorphism. Materials and Methods: DNA was extracted from the paraffin-embedded tissue of 16 consecutive cases of endoscopic biopsy, eight of which supplied both dysplastic and nondysplastic tissue for comparison. HUMARA was amplified by PCR with or without pretreatment with methylationsensitive restriction enzyme, HpaII. The amplification products were electrophoresed on polyacrylamide gel and silver-stained. Results: Among the 16 cases, 13 cases were informative and 3 cases noninformative. Of the 13 cases, one case showed skewed lyonization, rendering 12 cases to be analyzed further. A monoclonal band pattern was noted in 2 cases, and a polyclonal band pattern in 10 cases. A review of the histopathologies of the monoclonal and the polyclonal cases did not reveal features discriminating the two groups. Conclusion: These results suggest that gastric dysplasia is a disease entity heterogeneous in the genetic level, and many cases may be non-neoplastic.

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주행패턴의 상대 가속도에 따른 중소형 자동차의 온실가스 및 대기오염물질 배출 특성 (Greenhouse Gas and Pollutant Emission from Light-Duty Vehicles Regarding the Relative Positive Acceleration)

  • 이태우;길지훈;박경균;박준홍;박용희;홍지형;이대엽
    • 한국자동차공학회논문집
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    • 제18권4호
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    • pp.31-39
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    • 2010
  • Although driving patterns strongly influence greenhouse gas and air pollutant emission rate from light duty vehicles, emission measurements have been mainly based on chassis dynamometer testing with one standard driving pattern. And there has been limited work on quantifying the independent effect of driving parameters on emission rate because of multidimensional nature of real-world driving pattern. The objective of this study is to obtain the quantitative effect of relative positive acceleration (RPA) on vehicle emission rate. RPA has been used to define the occurrence of acceleration demanding large amounts of power in certain driving distance and shown to be a significant affecting parameter for real-world emission rate. 40 driving patterns have been developed with fixed driving parameters to investigate independent effect of RPA. For the same values of average vehicle speed and power, the trend in carbon dioxide emission rate and fuel consumption with respect to RPA is very clear. Emission rate of nitrogen oxide and particulate matter also increase with respect to RPA, but the trend is less clear. Carbon dioxide emission from diesel vehicle appear to be more affected by high accelerations compared to that from gasoline vehicle because of high intake air restriction during acceleration caused by turbocharger and intercooler. The results have implications for the possible reduction of environmental effects through better traffic planning and management, driver education and car design.

자가불화합성 Brassica campestris에 있어서 단일유전자좌가설에 의해 분리되지 않는 S-유전자 계통의 분석 (Analysis of Non-segregated S-allele Strain by Single-Locus Hypothesis in Self-incompatible Brassica campestris)

  • 노일섭
    • Journal of Plant Biology
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    • 제36권2호
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    • pp.127-132
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    • 1993
  • Self-incompatibility in Brassica campestris is controlled by multi-allele system in a single genetic locus, the S locus, and it is elucidated that S-glycoproteins are S gene products. In this experiments, we examined the genetic mode(pollen tube behavior and segregation of S-glycoprotein), characteristic of S-glycoproteins and DNA constitution within nuclear genome on S gene family that unexplained by single locus model, and investigated the segregation pattern of S-glycoproteins in bred F1 generation. By diallel cross among the 15 plants within one family the existence of three types of homozygotes and three types of heterozygotes were observed, and segregation of S-allele could not explained by single locus model. From the results of IEF-immunoblot analysis for non-segregated individual plant, the segregation pattern of S specific bands was corresponded with results of diallel cross except with one case(SaSa genotype). The molecular weight of 6 different S-genotype varied in near by 50 kD, and each genotype expressed with 2 or 3 bands. Specific bands in SaSa, SbSb, ScSc has almost similar molecular weight between them. Southern analysis of genomic DNA probed with S-glycoprotein cDNA for 6 different genotypes revealed that there are clear difference in polymorphism, multiple bands of hybridization, when restriction enzymes of EcoR I were used. It could be assumed that there are several sequences related to the S-glycoprotein structural genes within their nuclear genome. Therefore, we suggested the possibilities that S-allele system could be controlled by multi-locus, that dominance-recessive interactions could be explained by modifier gene or supressor gene based on the results of abnormal segregation of S-glycoprotein in bred F1. The F2 analyses are progressing in now.

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중년기 남녀의 체중 감량 시도 여부에 따른 건강 관련 생활습관과 식행동의 차이 (Relations Among Weight Control Behaviors, Health-related Lifestyles, and Diet Behaviors in Middle Aged Koreans)

  • 최윤정;김은미
    • 대한지역사회영양학회지
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    • 제13권2호
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    • pp.176-188
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    • 2008
  • In this study, we compared demographic anthropometric characteristic, health-related lifestyle and diet behavior among weight control behaviors of 1187 (555 male, 632 female) aged $40{\sim}69yrs$ in Ganghwa country. All the data were analyzed by chi-square test, trend test, student t-test using SPSS 12.0 version at p < 0.05. 'Attempting weight control (loss)' was more in women than that was found in men (36.6% vs 20.7%), and women attempting weight loss most were 40-50 yrs. The reasons of weight loss were 'health problem' and 'health promotion'. Physical activity and diet restriction were commonly employed as weight control methods. Both genders attempting weight loss had a higher education level, BMI, percentage of body fat, waist circumference and physical activity than those not attempting weight control (p < 0.05). In dietary habits like 'meal regularity', 'slow eating' and 'over eating', women attempting weight loss were superior than those who not attempting weight control group (p < 0.05). Eating pattern changes like 'decrease of fats and fatty foods intake', 'vegetable oil usage', 'increase of fruit and vegetables intake', 'decrease of sugar and salt intake' showed significant differences (p < 0.001) between the attempted weight control groups and nonattempted weight control groups. Salt taste was a preference in male non-attempted weight control group, while sour, hot and spicy taste were preference in female attempted weight control group (p < 0.05). Preference for processed foods, fried foods and snack were significant differences (p < 0.05) in women attempted weight control group. Those attempting weight loss tried to improve their eating patterns. However, those attempting weight loss were poorer than the others in health-related lifestyle and eating habit. Therefore, it is necessary to make an effort that improve healthrelated lifestyle and diet behavior in middle aged group.

The variation of insulin like growth factor 2 maker is associated with growth traits in Thai native (Kradon) pigs

  • Kessara Ampaporn;Rattikan Suwannasing;Pitchayanipa Phongphanich;Supanon Tunim;Monchai Duangjinda
    • Animal Bioscience
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    • 제36권9호
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    • pp.1350-1356
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    • 2023
  • Objective: This study was conducted to investigate polymorphisms of the melanocortin-4 receptor (MC4R) and insulin like growth factor 2 (IGF2) genes and to evaluate the growth traits affected by such polymorphisms in Thai native (Kradon) pigs. Methods: Blood samples and productive data from 91 Kradon pigs were collected. DNA was extracted and quantified, the IGF2 and MC4R genes were amplified, and the polymerase chain reaction (PCR) produces were digested using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique. Genotyping was performed, and the association between genotypes and growth traits on the birth and weaning weights were evaluated. Results: The IGF2 intron7 g.162G>C variations in Kradon pigs were found in three genotypes: i) GG, ii) GC, and iii) CC. The GG genotype frequency was the highest followed by the GC and CC genotypes. The frequencies of the G and C alleles were 0.703 and 0.297, respectively. The MC4R genotype was found in only one genotype (GG). The IGF2 gene pattern was not associated with birth weight traits, whereas the IGF2 gene pattern was related to the weaning weight trait in Kradon pigs. Pigs with the CC and GC genotypes had higher weaning weights than ones with the GG genotype (p<0.001). Conclusion: Thai native Kradon pigs with the CC and GC genotypes of the IGF2 gene have higher weaning weights than pigs with the GG genotype.

인터넷을 활용한 패턴 학습에서의 수학적 의사소통 및 문제해결에 관한 연구 (A Study on the Mathematical Communication and Problem Solving Using Internet in Pattern)

  • 류성림;박신정
    • 대한수학교육학회지:학교수학
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    • 제5권4호
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    • pp.459-476
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    • 2003
  • 본 연구는 수학 교실에서의 시·공간적 제약에서 벗어나 자유롭게 수학 문제를 해결하고 다양한 수학적 의사소통을 할 수 있는 환경을 조성하기 위하여 인터넷을 도입하였다. 인터넷을 활용한 수학 교육에서 나타나는 수학적 의사소통과 문제해결의 형태를 알아보았다. 본 연구를 통하여 인터넷 환경에서 수학적 아이디어를 표현하는 방법은 학생들마다 다양한 유형으로 나타났으며, 인터넷을 활용한 학습은 다른 학생들의 표현 방법을 통해 자신의 표현 방법을 보다 논리적으로 개선하는 데 도움을 줄 수 있는 것으로 보여진다. 학습자의 특성에 따라 수학적 의사소통 참여 도에 차이가 있었으며, 학업성취도가 낮은 학생들에게는 수학에 대한 흥미와 자신감을 갖게 하는데 도움이 되었다. 또한 인터넷을 활용한 패턴 학습이 수학 교실의 문제해결에 있어서 이유 및 검증의 습관화와 과제 집착력 증가에 영향을 미쳤다.

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