• Title/Summary/Keyword: RFLP.

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지방산 결합 단백질(FABP) 유전자를 이용한 한우 도체 및 육질 관련 SNP 분자 표지 개발

  • Sin, Seong-Cheol;Kim, Gi-Rak;Park, Jong-Geun;Sin, Gi-Hyeon;Lee, Jun-Je;Heo, Yeon-Beom;Jeong, Gu-Yong;Jeong, Eui-Ryong
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2006.05a
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    • pp.117-121
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    • 2006
  • 본 연구는 한우의 도체 품질을 결정하는 육질 등급 판정 항목이자 경제적으로 매우 중요한 근내지방도, 배최장근 단면적 및 등지방두께에 대한 개체별 유전능력 차이를 조기에 식별하는 선발 기술을 개발하기 위해 세포내 지방산 농도 및 다양한 세포 및 지질대사를 조절하는 지 방산결합 단백질(H-FABP) 유전자의 전체 염기서열을 분석하여 SNP를 검출하고, SNP marker가 한우의 도체 및 육질 형질에 미치는 영향에 대하여 분석하고자 수행하였다. 염기 서열 분석 결과 총 6개의 SNP를 검출하였고, 이들 중 4개의 주요 SNP를 선발하여 PCR-RFLP기법으로 genotyping하고, 각 SNP marker가 한우 도체 및 육질 형질에 미치는 영향을 통계분석 하였다. H-FABP의 C3523T SNP marker가 한우의 배최장근 단면적 및 등지방 두께에 유의적인 영향을 미쳤다(p<0.05). 따라서, 본 연구를 통해 개발된 한우 H-FABP 유전자의 특정 SNP marker는 배최장근 단면적은 넓고 등지방두께는 얇은 우수한 고급육을 생산하는 한우의 조기 식별 및 육질진단에 매우 유용한 SNP분자 표지로 활용할 수 있을 것으로 기대된다.

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Effect of Single Nucleotide Polymorphism of Endothelial Differentiation G-Protein Coupled Receptor 1 (EDG1) Gene on Marbling Score in Hanwoo

  • Shin, Sung-Chul;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.32 no.6
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    • pp.776-782
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    • 2012
  • Marbling (intramuscular fat) is the most economically important meat quality trait in Hanwoo (Korean cattle). The endothelial differentiation G-protein coupled receptor 1 (EDG1) gene, involved in blood vessel formation, is located within the genomic region of a quantitative trait locus (QTL) for marbling on bovine chromosome 3. Thus, the EDG1 gene can be considered as a positional and functional candidate gene for meat quality in beef cattle. This study aimed to identify single nucleotide polymorphisms (SNPs) in the EDG1 gene and to evaluate their associations with carcass traits in Hanwoo population. We have sequenced a fragment of 5'-UTR of the EDG1 gene and identified one SNP. Genotyping of the g.166A>G SNP marker was carried out using PCR-RFLP analysis in 309 Hanwoo steers in order to evaluate their association with carcass traits. The g.166A>G SNP marker showed a significant effect on the marbling score. Animals with the GG genotype had higher marbling score compared with AA and AG genotypes (p<0.05). This SNP marker also showed a significant additive effects for the marbling score (p<0.05). These results suggest that the EDG1 gene can be used as a molecular marker for DNA marker-assisted selection in order to increase the levels of the marbling score in Hanwoo.

Molecular Characterization of a New Hantaan Virus Howang Strain (한탄바이러스 호왕균주의 M, S 유전자 절편의 염기서열 및 분자생물학적 특성)

  • Chu, Yong-Kyu;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.59-68
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    • 1997
  • Hantaan virus Howang strain which isolated from the blood of severe case of Korean hemorrhagic fever is more virulent than HTN 76/118 and showed different RFLP from partial PCR amplifed M genome segment to established Hantaan serotype viruses. We have determined the nucleotide sequence of the M and S genome segments and compared to HTN 76/118. The M and S segment of Howang strain has 3615 and 1696 nucleotides long, respectively. The M segment sequence of Howang strain is one mucleotide shorter than HTN 76/118. The sequence data of Howang strain shows 93.5% homology to HTN 76/118. One long open reading frame, which strats from 41nt. to 3448nt. of the M segment and from 37nt. to 1326nt. of the S segment, exist to on complementary sense of the virus genome. There are no significant difference between HTN 76/118 and Howang strain on hydrophobicity of deduced polypeptides, but has slight difference on secondary structure.

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Construction of Molecular Genetic Linkage Map Using RAPD Markes in Cowpea

  • Chung, Jong-Il;Shim, Jung-Hyun;Go, Mi-Suk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.4
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    • pp.341-343
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    • 2001
  • Molecular markers have become fundamental tools for crop genome study. The objective of this study was to construct a genetic linkage map for cowpea with PCR-based molecular markers. Five hundred and twenty random RAPD primers were screened for parental polymorphism. Ninety RAPD markers from sixty primers was segregated in 75 F2 mapping population derived from the cross of local cultivars GSC01 and GSC02. 70 RAPD markers were found to be genetically linked and formed 11 linkage groups. Linkage map spanned 474.1 cM across all 11 linkage groups. There are six linkage groups of 40 cM or more, and five smaller linkage groups range from 4.9 to 24.8 cM. The average linkage distance between pairs of markers among all linkage groups was 6.87 cM. The number of markers per linkage group ranged from 2 to 32. The longest group 1 spans 190.6 cM, while the length of shortest group 11 is 4.9 cM. This map is further needed to be saturated with the various markers such as RFLP, AFLP, SSR and more various populations and primers. In addition, morphological markers and biochemical markers should be united to construct a comprehensive linkage map.

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Cloning and Characterization of Highly Repetitive Sequences in the Genome of Allium sativum L. (마늘(Allium sativum L.) 게놈의 고반복서열의 분이와 특성 조사)

  • 이동희
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.49-55
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    • 1996
  • We have studied the DNA of Allium sativum L. with respect to highly repetitive sequences. Fast reassociated DNA fragments expected to be highly repetitive sequences based on $C_{o}t$ curve were isolated and characterized. Their copy numbers were approximately $10^{5}~10^{7}$ per haploid genome. Nucleotide sequences analysis of six candidates reveals that their G/C content were low, 25-40% and typical patterns of repeating sequences exist. Repeat sequences were used as probes to access restriction fragment length polymorphism (RFLP) of genomic DNAs of four local clones, Tanyang, Mungyong, So san, and Uisong. The hybridization pattern were very similar among these four local clones.clones.

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Colonizing Ability of Pseudomonas fluorescens 2112, Among Collections of 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens spp. in Pea Rhizosphere

  • Kim, Sang-Dal;Fuente, Leonardo De La;Weller, David M.;Thomashow, Linda S.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.763-770
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    • 2012
  • Pseudomonas fluorescens 2112, isolated in Korea as an indigenous antagonistic bacteria, can produce 2,4-diacetylphloroglucinol (2,4-DAPG) and the siderophore pyoveridin2112 for the control of phytophthora blight of red-pepper. P. fluorescens 2112 was classified into a new genotype C among the 17 genotypes of 2,4-DAPG producers, by phlD restriction fragment length polymorphism (RFLP). The colonizing ability of P. fluorescens 2112 in pea rhizosphere was equal to the well-known pea colonizers, P. fluorescens Q8r1 (genotype D) and MVP1-4 (genotype P), after 6 cycling cultivations for 18 weeks. Four tested 2,4-DAPG-producing Pseudomonas spp. could colonize with about a 96% dominance ratio against total bacteria in pea rhizosphere. The strain P. fluorescens 2112 was as good a colonizer as other Pseudomonas spp. genotypes in pea plant growth-promoting rhizobacteria.

An Association Study of the Brain-Derived Neurotrophic Factor Val66Met Gene Polymorphism and Schizophrenia (Brain-Derived Neurotrophic Factor Val66Met 다형성과 정신분열병의 관련 연구)

  • Lee, Hwa-Young;Kim, Dae-Jin;Kim, Yong-Ku
    • Korean Journal of Biological Psychiatry
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    • v.13 no.4
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    • pp.267-272
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    • 2006
  • Objectives : Schizophrenia is a clinically heterogenous disease with a strong genetic component. Many studies have suggested that brain-derived neurotrophic factor(BDNF) is involved in the pathophysiology of schizophrenia. This study was performed to determine whether there is an association between BDNF Val66Met polymorphism and schizophrenia. Methods : To identify any genetic predisposition to schizophrenia, we investigated the BDNF Val66Met polymorphism in 106 patients with schizophrenia and 147 normal controls with PCR-RFLP method. Statistical analyses were used to test the association between and BDNF Val66Met genotype and Schizophrenia. Results : No association was found between BDNF Val66Met polymorphism and schizophrenia. No significant differences were found comparing the BDNF genotype distributions according to the age of onset, the number of admission and familial loading in schizophrenia. Conclusion : This result indicates that BDNF Val66Met polymorphism is not associated with schizophrenia. However, further studies with a large number of subjects are needed to confirm whether the BDNF gene is related to schizophrenia.

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Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
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    • v.3 no.1
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    • pp.73-78
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    • 1999
  • Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

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The Diversity of Culturable Organotrophic Bacteria from Local Solar Salterns

  • Yeon, Sun-Hee;Jeong, Won-Jin;Park, Jin-Sook
    • Journal of Microbiology
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    • v.43 no.1
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    • pp.1-10
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    • 2005
  • We isolated and cultured bacteria inhabiting solar saltern ponds in Taean-Gun, Chungnam Province, Korea. All of the isolated 64 strains were found to be moderately halophilic bacteria, growing in a salt range of 2-20 %, with an optimal concentration of 5% salt. Bacterial diversity among the isolated halophiles was evaluated via RFLP analyses of PCR-amplified 16S rDNAs, followed by phylogenetic analysis of the partial 16S rDNA sequences. The combination of restriction enzyme digestions with HaeIII, CfoI, MspI and RsaI generated 54 distinct patterns. A neighbor-joining tree of the partial 16S rDNA sequences resulted in the division of the 64 strains into 2 major groups, 45 strains of ${\gamma}-Proteobacteria$ (70.3%) and 19 strains of Firmicutes (29.7%). The ${\alpha}-Proteobacteria$ and Cytophaga-Flavobacterium-Bacterioides groups, which were repeatedly found to exist in thalassohaline environments, were not represented in our isolates. The ${\gamma}-Proteobacteria$ group consisted of several subgroups of the Vibrionaceae (37.5%), Pseudoalteromonadaceae (10.9%), Halomonadaceae (7.8%), Alteromonadaceae (7.8%), and Idiomarinaceae (6.3%). Members of Salinivibrio costicola (29.7%) were the most predominant species among all of the isolates, followed by Halobacillus treperi (12.5%). Additionally, three new species candidates were found, based on similarities of the 16S rDNA sequences to those of previously published species.

Identification Based on Computational Analysis of rpoB Sequence of Bacillus anthracis and Closely Related Species (Bacillus anthracis와 그 유연종의 rpoB 유전자 컴퓨터 분석을 통한 동정)

  • Kim, Kyu-Kwang;Kim, Han-Bok
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.333-338
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    • 2008
  • Computational analysis of partial rpoB gene sequence (777 bp) was done in this study to identify B. anthracis and its closely related species B. cereus and B. thuringiensis. Sequence data including 17 B. anthracis strains, 9 B. cereus strains, and 7 B. thuringiensis strains were obtained by searching databases. Those sequences were aligned and used for other computational analysis. B. anthracis strains were identificated by in silico restriction enzyme digestion. B. cereus and B. thuringiensis were not segregated by this method. Those sequencing and BLAST search were required to distinguish the two. In actual identification tests, B. anthracis strains could be identified by PCR-RFLP, and B. cereus and B. thuringiensis strains were distinguished by BLAST search with reliable e-value. In this study fast and accurate method for identifying three Bacillus species, and flow chart of identification were developed.