• Title/Summary/Keyword: Nucleotide Sequence

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Genetic Variation and Phylogenetic Relationship of Taraxacum Based on Chloroplast DNA (trnL-trnF and rps16-trnK) Sequences (엽록체 DNA (trnL-trnF, rps16-trnK) 염기서열에 의한 국내 민들레속 유전자원의 유전적 변이와 유연관계 분석)

  • Ryu, Jaihyunk;Lyu, Jae-il;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.30 no.5
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    • pp.522-534
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    • 2017
  • This study was investigated genetic variation in 24 Taraxacum accessions from various regions in South Korea based on the sequences of two chloroplast DNA (cpDNA) regions (trnL-trnF and rps16-trnK). T. mongolicum, T. officinale, and T. laevigatum were triploid, and T. coreanum and T. coreanum var. flavescens were tetraploid. The trnL-trnF region in native Korean dandelions (T. mongolicum, T. coreanum, and T. coreanum var. flavescens) were ranged from 931 to 935 bp in length, and that of naturalized dandelions were ranged from 910 bp (T. officinale) to 975 bp (T. laevigatum) in length. The rps16-trnK region in T. mongolicum, T. coreanum, T. coreanum var. flavescens, T. officinale, and T. laevigatum was 882-883 bp, 875-881 bp, 878-883 bp, 874-876 bp, and 847-876 bp, respectively, in length. The sequence similarity matrix of the trnL-trnF region ranged from 0.860 to 1.00 with an average of 0.949, and that of the rps16-trnK region ranged from 0.919 to 1.000 with an average of 0.967. According to the phylogenetic analysis, the Korean native taxa and naturalized taxa were divided independent clade in two cpDNA region. T. coreanum var. flavescens clustered only with T. coreanum, and there were no significant differences in their nucleotide sequences. The finding that two accessions (T. coreanum; Jogesan, T. mongolicum; Gangyang) had a high level of genetic variation suggests their utility for breeding materials.

The Differentially Expressed Genes by Radiotherapy in the Patients with Uterine Cervix Cancer (자궁경부암 환자에서 방사선치료 시 발현되는 유전자의 규명)

  • Seo Eun Young;Cho Moon-June;Lee Jeung Hoon;Lee Young-Sook;Na Myung-Hoon;Lee Woong-Hee;Kim Jun-Sang;Kim Jae-Sung
    • Radiation Oncology Journal
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    • v.19 no.4
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    • pp.389-396
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    • 2001
  • Purpose : To detect differentially expressed genes in the patients with uterine cervical cancer during the radiation therapy. Materials and Methods : In patients with biopsy proven uterine cervical cancer, we took tumor tissue just before radiation therapy and at 40 minutes after external irradiation of 1.8 Gy. Total RNAs isolated from non-irradiated and irradiated tumor tissue samples were analyzed using the differential-display reverse transcription-polymerase chain reaction (DDRT-PCR). Complementary DNA (cDNA) fragments corresponding to differentially expressed messenger RNAs(mRNAs) were eluted, and cloned. The differential expression of the corresponding mRNAs was confirmed by reverse northern blot. Differentially expressed cDNA bands were sequenced. Nucleotide sequence data were analyzed in the Gene Bank and EMBL databases via the BLAST network sewer to identify homologies to known genes or cDNA fragments. Expression pattern of down-regulated clone was examined using RT-PCR in S patients undergoing radiotherapy. Results : We identified 18 differentially expressed bands by DDRT-PCR, which were eluted and cloned. There were 10 up-regulated clones and 1 down-regulated clone in reverse northern blot. One cDNA fragment had homology to chemokine receptor CXCR4, four were identified as Human ESTs in the EMBL database in EST clones. Down-regulated CxCa-11 was also down regulated in all patients. Conclusion : Using the DDRT-PCR, we have identified 10 up-regulated and 1 down-regulated clone(s) in the patients with uterine cervical cancer during the radiation therapy. The clinical relevance and the functions of these genes will be further investigated.

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Effects of SNP Markers of the Apolipoprotein E (APOE) Gene on Meat Quantity and Quality Traits in Korean Cattle (한우 아포지단백질 E (APOE) 유전자의 SNP Marker가 육량 및 육질형질에 미치는 영향)

  • Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.108-113
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    • 2009
  • Apolipoprotein E (APOE) is a plasma lipoprotein in mammals and plays an important role in the transport and metabolism of lipids such as phospholipids and triglycerides. Therefore, the APOE gene could be a candidate gene controlling lipid metabolism in beef cattle. This study was performed to identify single nucleotide polymorphisms (SNP) in the APOE gene and to investigate the effects of SNP genotype on the carcass traits such as meat quantity and quality in Korean cattle. For PCR amplification, pooled DNA made from unrelated 60 individuals was prepared and primer pairs were designed based on the cDNA sequence of exon 4 region of the bovine APOE gene. A SNP was identified at position 2034 (T/C substitution) of the exon 4 region in the APOE gene. PCR-RFLP procedure with restriction enzyme ACC I was developed for determining the SNP genotype for each of a total of 309 animals with pedigree information and performance records through the national progeny testing program. The frequencies of the genotypes TT, TC and CC were 10.9, 46.9 and 42.2%. Gene frequencies were 0.344 for T allele and 0.656 for C allele. The g.2034T>C SNP genotype showed a significant effect (p<0.05) on dressing percentage and meat color, respectively. Animals with the TT genotype showed higher dressing percentage than those with the CC genotype, and TT genotype had desirable meat color compared with CC genotype. These results suggest that the g.2034T>C SNP genotype of the APOE gene may be useful as a DNA marker for meat quantity index and dressing percentage in Korean cattle.

Molecular Characterization of Cinnamate 4-Hydroxylase gene in Red Hot Pepper (Capsicum annuum L.) (고추에서 분리한 Cinnamate 4-Hydroxylase 유전자의 분자생물학적 특성)

  • Kim Kye-Won;Ha Sun-Hwa;Cho Kang-Jin;Kim Eun-Ju;Lee Min-Kyung;Yu Jae-Ju;Kim Jong-Guk;Lee Shin-Woo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.167-173
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    • 2005
  • Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.

Intracullular Functions of the mas2+ Gene in the Fission Yeast, Schizosaccharomyces pombe (분열형 효모에서의 mas2+ 유전자의 세포 내 기능)

  • Sin, Sang-Min;Cha, Jae-Young;Ha, Se-Eun;Sim, Sun-Mi;Kim, Hyoung-Do;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.19 no.1
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    • pp.101-110
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    • 2009
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel $mas2^+$ (mitosis associated protein) gene, a homolog of human SMARCAD1 was isolated and characterized from a fission yeast Schizosaccharomyces pombe (S. pombe) using gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 922 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that an SNF2 domain is located, which is involved in the chromosome remodeling. The quantitative analysis of the $mas2^+$ transcript against $adh1^+$ showed that the expression level of $mas2^+$ is high before septum formation in S. pombe. When $mas2^+$ null mutant cells were grown at 27 and $35^{\circ}C$, the cytokinesis of $mas2^+$ null mutant was greatly delayed and a large number of multi-septate and mis-segregated cells were produced. In addition, the number of multi-septate cells significantly increased. When cells were cultured in YES rich medium to increase proliferation, the abnormal phenotypes $mas2^+$ null mutant dramatically increased. These phenotypes could be rescued by an over-expression of the mast gene. The Mas2 protein localized in the nuclei of S. pombe, as evidenced by Mas2-EGFP signals. These results suggest that the $mas2^+$ is homologous to human SMARCAD1 gene and involved in septum formation and chromosome remodeling control.

Association Between the Polymorphism on Intron 5 of the Lipoprotein Lipase Gene and Carcass Traits in Hanwoo (Korean cattle) (한우 Lipoprotein Lipase 유전자 Intron 5번의 Polymorphism과 경제 형질과의 관련성 분석)

  • Lee, H.J.;Lee, S.H.;Cho, Y.M.;Yoon, H.B.;Jeon, B. K.;Oh, S.J.;Kwon, M.S.;Yoon, D.H.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.947-956
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    • 2004
  • The primary role of lipoprotein lipase(LPL) is the hydrolysis of triglycerides(TG) from the core of triglyceride-rich lipoproteins such as chylomicrons and very low density lipoproteins in plasma. Fatty acids liberated by LPL on capillary endothelial surfaces are available for tissues as energy sources especially in muscles or for storage in the form of TG in adipose tissues. Therefore, as the candidate gene related to the carcass traits of the beef cattle, we have directly sequenced the exon 5${\sim}$exon 6 region in the bovine LPL gene for discovery of single nucleotide polymorphism(SNP) with 24 unrelated Hanwoo(Korean cattle). Novel eight sequence variants were detected: three loci on exon 5, three on intron 5 and two on exon 6. All SNPs identified were strongly linked each other, and one hundred twenty eight Hanwoo samples were genotyped one SNP on intron 5 using PCR-restriction fragment length polymorphism method by digestion with Hae III restriction enzyme. The allele frequency of the polymorphism was 0.76 and 0.24. The effects of this polymorphism on the breeding values of the carcass weight, loin muscle area, back fat thickness and marbling score were analyzed using least square methods of SAS GLM. The marbling score of BB genotype was significantly higher than those of AA and AB genotypes(P<0.05). This result indicates that this polymorphism may be associated with the variation of marbling score. Further study is warranted to investigate the phenotypic association in Hanwoo.

Single Nucleotide Polymorphisms (SNPs) Discovery in GHSR Gene and Their Association Analysis with Economic Traits in Korean Native Chickens (GHSR 유전자 내 유전변이의 탐색과 한국재래계의 성장 및 산란 특성에 미치는 연관성 분석)

  • Choi, So-Young;Hong, Min-Wook;Yang, Song-Yi;Kim, Chong-Dae;Jeong, Dong Kee;Hong, Yeong Ho;Lee, Sung-Jin
    • Korean Journal of Poultry Science
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    • v.43 no.4
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    • pp.273-279
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    • 2016
  • Recently, it was reported that certain polymorphisms in the growth hormone secretagogue receptor gene (GHSR) are associated with the growth of chickens. However, the correlation between GHSR polymorphisms and economic traits has not been investigated in Korean native chickens (KNCs). Therefore, the objective of this study was to confirm the suitability of the GHSR gene as a candidate for genomic selection and identify a genetic marker for KNCs. A total of 220 KNCs from six breeds raised at the National Institute of Animal Science were genotyped for the c.739+726 SNP in the GHSR gene using polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP), and the sequence for a subset of 30 birds was analyzed using direct sequencing. The association between the SNP genotypes and the economic traits of the KNCs was analyzed using the statistical package for the social science (SPSS) software program. The association analysis between the c.739+726T>C SNP and economic traits revealed that the SNP was significantly associated with body weight at 150 and 270 days (BW150 and BW270, respectively) in all KNCs (p<0.01), BW150 in KNC (Gary) (p<0.05), and egg production number in KNC (White, p<0.05). In addition, the SNPs discovered using direct sequencing (513A>G, 517A>T) had a significant effect on the body weight and egg production traits (p<0.05). In conclusion, these results might be useful as a basis for studies on the improvement of KNC breeds. Furthermore, these results suggest that the SNPs (c.739+726T>C, 513A>G, and 517A>T) located in the GHSR gene could be useful molecular genetic markers for KNCs.

Current Status of Cattle Genome Sequencing and Analysis using Next Generation Sequencing (차세대유전체해독 기법을 이용한 소 유전체 해독 연구현황)

  • Choi, Jung-Woo;Chai, Han-Ha;Yu, Dayeong;Lee, Kyung-Tai;Cho, Yong-Min;Lim, Dajeong
    • Journal of Life Science
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    • v.25 no.3
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    • pp.349-356
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    • 2015
  • Thanks to recent advances in next-generation sequencing (NGS) technology, diverse livestock species have been dissected at the genome-wide sequence level. As for cattle, there are currently four Korean indigenous breeds registered with the Domestic Animal Diversity Information System of the Food and Agricultural Organization of the United Nations: Hanwoo, Chikso, Heugu, and Jeju Heugu. These native genetic resources were recently whole-genome resequenced using various NGS technologies, providing enormous single nucleotide polymorphism information across the genomes. The NGS application further provided biological such that Korean native cattle are genetically distant from some cattle breeds of European origins. In addition, the NGS technology was successfully applied to detect structural variations, particularly copy number variations that were usually difficult to identify at the genome-wide level with reasonable accuracy. Despite the success, those recent studies also showed an inherent limitation in sequencing only a representative individual of each breed. To elucidate the biological implications of the sequenced data, further confirmatory studies should be followed by sequencing or validating the population of each breed. Because NGS sequencing prices have consistently dropped, various population genomic theories can now be applied to the sequencing data obtained from the population of each breed of interest. There are still few such population studies available for the Korean native cattle breeds, but this situation will soon be improved with the recent initiative for NGS sequencing of diverse native livestock resources, including the Korean native cattle breeds.

Microsatellite Alterations of Plasma DNA in Non Small Cell Lung Cancer (비소세포폐암 환자의 혈장 DNA를 이용한 Microsatellite 분석)

  • Kim, Kyu-Sik;Kim, Eun-Jung;Kim, Soo-Ock;Oh, In-Jae;Park, Chang-Min;Jeong, Ju-Yeon;Kim, Yu-Il;Lim, Sung-Chul;Park, Jong-Tae;Kim, Young-Chul
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.4
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    • pp.352-358
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    • 2005
  • Microsatellites are short tandem repeated nucleotide sequences that are present throughout the human genome. Variations in the repeat number or a loss of heterozygosity around the microsatellites have been termed a microsatellite alteration (MA). A MA reflects the genetic instability caused by an impairment in the DNA mismatch repair system and is suggested to be a novel tumorigenic mechanism. A number of studies have reported that MA in the DNA extracted from the plasma occurs at varying frequencies among patients with a non-small cell lung carcinoma (NSCLC). The genomic DNA from 9 subjects with a non-small cell lung cancer (squamous cell cancer 6, adenocarcinoma 2, non-small cell lung cancer1) and 9 age matched non-cancer control subjects (AMC: tuberculosis 3, other inflammatory lung disease 6) and 12 normal control subjects (NC) were extracted from the peripheral blood leukocytes and plasma. Three microsatellite loci were amplified with the primers targeting the Gene Bank sequence D21S1245, D3S1300, and D3S1234. MA in the form of an allelic loss or a band shift was examined with 6% polyacrylamide gel electrophoresis and silver staining. None (0/12) of the NC subjects less than 40 years of age showed a MA in any of the three markers, while 88.9%(8/9) of the AMC above 40 showed a MA in at least one of the three markers (p<0.05). Sixty percent(6/10) of the control subjects with a smoking history showed a MA in one of the three markers, while 9.1%(1/11) of the control subjects without smoking history showed a MA (p<0.05). However, not only did 66.7%(6/9) of lung cancer patients show a MA in at least one of the three markers but so did 88.9%(8/21) of the AMC patients (p>0.05). In conclusion, a MA in the D21S1245, D3S1300, and D3S1234 loci using DNA extracted from the plasma was detected in 66.7% of lung cancer while no MA was found in the young non-smoking control subjects. However, many of the non-cancer control subjects (aged smokers) also showed a MA, which compromised the specificity of the MA analysis as a screening test. Therefore, a further study with a larger sample size will be needed.

α-Glucosidase inhibitory activity and protease characteristics produced by Bacillus amyloliquefaciens (Bacillus amyloliquefaciens로부터 생산된 protease 특성 및 α-glucosidase 저해활성)

  • Lee, Rea-Hyun;Yang, Su-Jin;Hwang, Tae-Young;Chung, Shin-Kyo;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.727-734
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    • 2015
  • In this study, three GRAS (generally recognized as safety) strain was isolated from Doenjang and Cheonggukjang and identified as a protease-producing microorganism, following the appearance of a clear zone around its colony when cultured on a medium containing skim milk. Based on an analysis of the nucleotide sequence of 16S ribosomal RNA, the strains wereas identified as Bacillus amyloliquefaciens and wereas therefore named Bacillus amyloliquefaciens CDD5, Bacillus amyloliquefaciens CPD4, and Bacillus amyloliquefaciens CGD3. Here, we analyzed the protease and ${\alpha}$-glucosidase inhibitory activities of the three B. amyloliquefaciens strains. Among the isolated strains, B. amyloliquefaciens CGD3 exhibited the highest protease activity (9.21 U/mL, 24 hr). The protease activities of B. amyloliquefaciens CDD5 and B. amyloliquefaciens CPD4 reached 1.14 U/mL and 8.02 U/mL, respectively, at 48 hr. The proteases from the three B. amyloliquefaciens strains showed the highest activities within a pH range of 8.0-9.0 at $50^{\circ}C$, and casein was found to be the preferred substrate on evaluating enzyme activity in the substrate specificity assay. The B. amyloliquefaciens strains exhibited maximal growth when the nutrient broth medium had an initial pH within the range of 5.0-10.0, 6-9% sodium chloride (NaCl), and 5% glucose. B. amyloliquefaciens CDD5 exhibited a low ${\alpha}$-glucosidase inhibition rate (5.32%), whereas B. amyloliquefaciens CPD4 and B. amyloliquefaciens CGD3 exhibited relatively higher inhibition rates of 96.89% and 97.55%, respectively.