• Title/Summary/Keyword: Direct Sequencing

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Discovery and Evaluation of Polymorphisms in the AKT2 and AKT3 Promoter Regions for Risk of Korean Lung Cancer

  • Sung, Jae-Sook;Park, Kyong-Hwa;Kim, Seung-Tae;Kim, Yeul-Hong
    • Genomics & Informatics
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    • v.10 no.3
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    • pp.167-174
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    • 2012
  • AKT is a signal transduction protein that plays a central role in the tumorigenesis. There are 3 mammalian isoforms of this serine/threonine protein kinase-AKT1, AKT2, and AKT3-showing a broad tissue distribution. We first discovered 2 novel polymorphisms (AKT2 -9826 C/G and AKT3 -811 A/G), and we confirmed 6 known polymorphisms (AKT2 -9473 C/T, AKT2 -9151 C/T, AKT2 -9025 C/T, AKT2 -8618G/A, AKT3 -675 A/-, and AKT3 -244 C/T) of the AKT2 and AKT3 promoter region in 24 blood samples of Korean lung cancer patients using direct sequencing. To evaluate the role of AKT2 and AKT3 polymorphisms in the risk of Korean lung cancer, genotypes of the AKT2 and AKT3 polymorphisms (AKT2 -9826 C/G, AKT2 -9473 C/T, AKT2 -9151 C/T, AKT2 -9025 C/T, AKT2 -8618G/A, and AKT3 -675 A/-) were determined in 360 lung cancer patients and 360 normal controls. Statistical analyses revealed that the genotypes and haplotypes in the AKT2 and AKT3 promoter regions were not significantly associated with the risk of lung cancer in the Korean population. These results suggest that polymorphisms of the AKT2 and AKT3 promoter regions do not contribute to the genetic susceptibility to lung cancer in the Korean population.

Conjugated linoleic acid producing potential of lactobacilli isolated from goat (AXB) rumen fluid samples

  • Tyagi, Amrish Kumar;Kumar, Sachin;Choudhury, Prasanta Kumar;Tyagi, Bhawna;Tyagi, Nitin
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.8
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    • pp.1233-1241
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    • 2020
  • Objective: The present investigation was aimed to explore the potential of lactobacilli for conjugated linoleic acid (CLA) production, isolated from rumen fluid samples of lactating goats. Methods: A total of 64 isolates of lactobacilli were obtained using deMan-Rogosa-Sharpe (MRS) agar from rumen fluid of goats and further subjected to morphological and biochemical characterizations. Isolates found as gram-positive, catalase negative rods were presumptively identified as Lactobacillus species and further confirmed by genus specific polymerase chain reaction (PCR). The phylogenetic tree was constructed from the nucleotide sequences using MEGA6. Results: Out of the 64 isolates, 23 isolates were observed positive for CLA production by linoleate isomerase gene-based amplification and quantitatively by UV-spectrophotometric assay for the conversion of linoleic acid to CLA as well as gas chromatography-based assay. In all Lactobacillus species cis9, trans11 isomer was observed as the most predominant CLA isomer. These positive isolates were identified by 16S rRNA gene-based PCR sequencing and identified to be different species of L. ingluviei (2), L.salivarius (2), L. curvatus (15), and L. sakei (4). Conclusion: The findings of the present study concluded that lactic acid bacteria isolated from ruminal fluid samples of goat have the potential to produce bioactive CLA and may be applied as a direct fed microbial to enhance the nutraceutical value of animal food products.

Association Study of Fibroblast Growth Factor 2 and Fibroblast Growth Factor Receptors Gene Polymorphism in Korean Ossification of the Posterior Longitudinal Ligament Patients

  • Jun, Jae-Kyun;Kim, Sung-Min
    • Journal of Korean Neurosurgical Society
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    • v.52 no.1
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    • pp.7-13
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    • 2012
  • Objective : The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of fibroblast growth factor (FGF) 2 gene and fibroblast growth factor receptor (FGFR) genes are associated with ossification of the posterior longitudinal ligament (OPLL). Methods : A total of 157 patients with OPLL and 222 controls were recruited for a case control association study investigating the relationship between SNPs of FGF2, FGFR1, FGFR2 and OPLL. To identify the association among polymorphisms of FGF2 gene, FGFR1, FGFR2 genes and OPLL, the authors genotyped 9 SNPs of the genes (FGF2 : rs1476217, rs308395, rs308397, and rs3747676; FGFR1 : rs13317 and rs2467531; FGFR2 : rs755793, rs1047100, and rs3135831) using direct sequencing method. SNPs data were analyzed using the SNPStats, SNPAnalyzer, Haploview, and Helixtree programs. Results : Of the SNPs, a SNP (rs13317) in FGFR1 was significantly associated with the susceptibility of OPLL in the codominant (odds ratio=1.35, 95% confidence interval=1.01-1.81, p=0.048) and recessive model (odds ratio=2.00, 95% confidence interval=1.11-3.59, p=0.020). The analysis adjusted for associated condition showed that the SNP of rs1476217 (p=0.03), rs3747676 (p=0.01) polymorphisms in the FGF2 were associated with diffuse idiopathic skeletal hyperostosis (DISH) and rs1476217 (p=0.01) in the FGF2 was associated with ossification of the ligament flavum (OLF). Conclusion : The results of the present study revealed that an FGFR1 SNP was significantly associated with OPLL and that a SNP in FGF2 was associated with conditions that were comorbid with OPLL (DISH and OLF).

Purification, Partial Characterization, and Immunoassay of Vitellogenin from Marbled Sole (Limanda yokohamae)

  • Kim Dae-Jung;Jung Jee-Hyun;An Cheul-Min;Jee Young-Ju;Min Kwang-Sik;Kim Yoon;Han Chang-Hee
    • Fisheries and Aquatic Sciences
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    • v.8 no.4
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    • pp.213-219
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    • 2005
  • Vitellogenin (VTG) was purified from the blood plasma of estradiol-17$\beta$ ($E_2$)-treated male marbled sole (Limanda yokohamae) using gel filtration and anion exchange chromatography. The purity of the marbled sole VTG (msVTG) was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE) and N-terminal amino acid sequencing. The purified msVTG was used to produce monoclonal and polyclonal antibodies in mice and rabbits, respectively, and the specificity of the polyclonal antisera for msVTG was confirmed by Western blot analysis. The antibodies cross­reacted with a protein of molecular mass approximately 160 kDa in the plasma samples of mature female marbled sole. No cross-reactivity was observed with the plasma of male fish. A direct non-competitive sandwich enzyme-linked immunosorbent assay (ELISA) was developed using the monoclonal anti-msVTG and polyclonal anti-msVTG antibodies, with purified msVTG as the standard protein. The values of the intra- and inter-assay variations were within the ranges of $8.l-9.8\%$ and $8.5-12.2\%$, respectively. The sensitivity was about 0.3 ng/mL. Serial dilutions of plasma from mature female sole reacted with the msVTG-antibodies in the sandwich ELISA, whereas the plasma from male fish did not. The results indicate that the maturation status of female marbled sole can be identified using a sandwich ELISA for msVTG.

Study on identification of candidate DNA marker related with beef quailty in QTL region of BTA 2 in Hanwoo population (한우 2번 염색체 양적형질좌위 영역에서 육질 연관 후보 DNA 마커 규명에 관한 연구)

  • Lee, Yoon-Seok;Oh, Dong-Yep;Yeo, Jung-Sou
    • Journal of the Korean Data and Information Science Society
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    • v.22 no.4
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    • pp.661-669
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    • 2011
  • By direct sequencing of 12 STS marker, we identified 10 polymorphic SNPs. As a result of genotype frequency analysis between 10 polymorphic SNPs and extreme population (n=20) for marbling score in Hanwoo (n=233), there was over 40 percent of frequency difference of HWSNP_1-1 and HWSNP_9-4 SNP. HWSNP_1-1 SNP was significantly associated with marbling score in large-scale population (n=233). Therefore we suggested that HWSNP_1-1 SNP can be useful as a positional candidate for beef quality for marker-assisted selection in Hanwoo.

False-Positive Mycobacterium tuberculosis Detection: Ways to Prevent Cross-Contamination

  • Asgharzadeh, Mohammad;Ozma, Mahdi Asghari;Rashedi, Jalil;Poor, Behroz Mahdavi;Agharzadeh, Vahid;Vegari, Ali;Shokouhi, Behrooz;Ganbarov, Khudaverdi;Ghalehlou, Nima Najafi;Leylabadlo, Hamed Ebrahmzadeh;Kafil, Hossein Samadi
    • Tuberculosis and Respiratory Diseases
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    • v.83 no.3
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    • pp.211-217
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    • 2020
  • The gold standard method for diagnosis of tuberculosis is the isolation of Mycobacterium tuberculosis through culture, but there is a probability of cross-contamination in simultaneous cultures of samples causing false-positives. This can result in delayed treatment of the underlying disease and drug side effects. In this paper, we reviewed studies on false-positive cultures of M. tuberculosis. Rate of occurrence, effective factors, and extent of false-positives were analyzed. Ways to identify and reduce the false-positives and management of them are critical for all laboratories. In most cases, false-positive is occurring in cases with only one positive culture but negative direct smear. The three most crucial factors in this regard are inappropriate technician function, contamination of reagents, and aerosol production. Thus, to reduce false-positives, good laboratory practice, as well as use of whole-genome sequencing or genotyping of all positive culture samples with a robust, extra pure method and rapid response, are essential for minimizing the rate of false-positives. Indeed, molecular approaches and epidemiological surveillance can provide a valuable tool besides culture to identify possible false positives.

Genetic polymorphism in regulatory region of fatty acid binding protein 4 (FABP4) and its effect on carcass weight in Hanwoo steers (한우 지방산결합단백질 4(FABP4) 유전자 조절영역내 단일염기변이(SNP)와 도체형질간 연관성 분석)

  • Lee, Seung-Hwan;Kim, Nam-Kuk;Kim, Seung-Chang;Choi, Bong-Hwan;Heo, Kang-Neung;Lee, Chang-Soo;Kim, Oun-Hyun;Lee, Jun-Heon;Kim, Hyeong-Cheul;Hong, Seong-Koo
    • Korean Journal of Agricultural Science
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    • v.38 no.4
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    • pp.673-680
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    • 2011
  • The aim of this study was to identify the polymorphism on fatty acid binding protein (FABP4) gene promoter region and its association with carcass traits in Hanwoo. We performed PCR-direct sequencing of FABP4 promoter region to identify single nucleotide polymorphism (SNPs) using unrelated 24 Hanwoo bulls. Four SNPs (-298A>G, -472A>G, -887A>G, -862A>G) were detected in the promoter region and genotyped on 583 Hanwoo steers. A linear mixed model revealed an association of three SNPs (-298A>G, -472A>G and -862A>G) with carcass weight and marbling score in dominance model (P<0.05). The animals with AA genotypes for the three SNPs were heavier carcass weight (5 kg) than animals with GG genotypes in the statistical analysis. For the marbling score, the AA genotype was lower effect of marbling score (0.21) than GG genotypes. In conclusion, this study indicates an important role for three SNPs detected in promoter region of FABP4 in determining carcass weight and marbling score in Hanwoo.

ADFP promoter polymorphism associated with marbling score in Korean cattle

  • Cheong, Hyun-Sub;Yoon, Du-Hak;Bae, Joon-Seol;Kim, Lyoung-Hyo;Kim, Eun-Mi;Kim, Ji-On;Hong, Jin;Kim, Nae-Soo;Shin, Hyoung-Doo
    • BMB Reports
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    • v.42 no.8
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    • pp.529-534
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    • 2009
  • Marbling score (MS) is the major trait that affects carcass quality in beef cattle. In this study, we investigated the association between genetic polymorphisms of the adipose differentiation-related protein gene (ADFP) and carcass traits in Korean cattle (also known as Hanwoo). Using direct DNA sequencing in 24 unrelated Korean cattle, 25 novel polymorphisms were identified within all exons and their flanking regions of ADFP, including the promoter region (1.5 kb). Among them, 21 polymorphic sites were selected for genotyping in the beef cattle (n = 425). Statistical analyses revealed that one promoter polymorphism (c.-56-18A > G) was associated with MS (P = 0.009). The 'A' allele of c.-56-18A > G exerted a lowering effect on MS, e.g., the lowest MS was found in 'A/A' (MS = 2.09 ${\pm}$ 1.23), intermediate in 'A/G' (MS = 2.11 ${\pm}$ 1.31), and the highest in 'G/G' (MS = 2.47 ${\pm}$ 1.47). Our findings suggest that these polymorphisms in ADFP might be important genetic factors involved in carcass quality in beef cattle.

Genetic Variation of the Mitochondrial Cytochrome b Sequence in Korean Rana rugosa (Amphibia; Ranidae)

  • Hyun Ick Lee;Dong Eun Yang;Yu Ri Kim;Hyuk Lee;Jung Eun Lee;Suh Yung Yang;Hei Yung Lee
    • Animal cells and systems
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    • v.3 no.1
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    • pp.89-96
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    • 1999
  • Nucleotide sequences of a 501 base-pair (bp) fragment in the mitochondrial cytochrome b (cyt b) gene were analyzed for 12 populations of Rana rugosa from Korea and Japan using a polymerase chain reaction (PCR) and direct silver sequencing. Two genetically distinct groups (type-A and type-B) were found in Korea. Type-A was found throughout most of South Korea and type-B was restricted to the mid-southeastern regions (Samchok, Yongdok, Chongsong and Pohang). But in the Tonghae population, both types were found. The level of mitochondrial DNA (mtDNA) sequence differences ranged from 0% to .0.8% among six populations of type-A, and 0 to 1.0% among 4 populations of type-B. However, sequence differences between type-A and type-B ranged from 5.4% to 6.6%, Using Kimura's two-parameter distance, the level of genetic sequence divergence between type-A and type-B was 6.7%. The Japanese R. rugosa was clustered very far from the Korean R. rugosa with 14.7%. In the neighbor-joining and UPGMA tree, all Korean samples were grouped, but subdivided into two types in 99% of the bootstrap iteration.

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A genetic algorithm for determining the optimal operating policies in an integrated- automated manufacturing system (통합자동생산시스템에서 최적운영방안 결정을 위한 유전자 알고리즘의 개발)

  • 임준목
    • Journal of Korea Society of Industrial Information Systems
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    • v.4 no.2
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    • pp.62-72
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    • 1999
  • We consider a Direct Input Output Manufacturing System(DIOMS) which has a munber of machine centers placed along a built-in Automated Storage/Retrieval System(AS/RS). The Storage/Retrieval(S/R) machine handles parts placed on pallets for the machine centers located at either one or both sides of the AS/RS. This paper deals with the operational aspect of DIOMS and determines the optimal operating policy by combining computer simulation and genetic algorithm. The operational problem includes: input sequencing control, dispatching rule of the S/R machine, machine center-based part type selection rule and storage assignment policy. For each operating policy, several different policies are considered based on the known research results. In this paper, using the computer simulation and genetic algorithm we suggest a method which gives the optimal configuration of operating policies within reasonable computation time.

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