• 제목/요약/키워드: rDNA sequencing

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천일염 생산공정별 미생물 분포 조사 및 호염미생물 동정 (Distribution and Identification of Halophilic Bacteria in Solar Salts Produced during Entire Manufacturing Process)

  • 나종민;강민승;김진효;김영섭;제정환;김정봉;조영숙;김재현;김소영
    • 한국미생물·생명공학회지
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    • 제39권2호
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    • pp.133-139
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    • 2011
  • 우리나라 전남지역에서 생산되는 천일염의 생산공정별 미생물 분포 조사 및 배양 분리법을 이용하여 호염미생물을 분리하여 동정하는 것을 이번 연구의 목표로 삼았다. 천일염 시료는 생산지를 고려하여 육지 염전인 영광군 한 지역과 해안 염전 지역인 신안군 두 군데에서 생산공정별로 세분화하여 총 28개 분석시료를 수집하여 사용하였으며, 이들 시료를 십진 희석하여 4종류의 미생물 분리용 배지에 도말, 배양한 후 나타난 집락(colony)을 계수하였다. 그 결과 천일염 생산 공정 중 대장균 등의 식품 오염지표 미생물은 검출되지 않았지만, 저장수에서 증발지 단계로 진행되면서 $1.1{\times}10^3{\sim}1.8{\times}10^5$ CFU/g의 호염성 미생물들이 검출되었다. 그러나 이후 단계인 함수저장고, 결정지에서는 미생물 수가 점차적으로 감소되었으며, 소금저장소에 보관된 천일염 시료에서는 미생물이 전혀 검출되지 않았다. 이들 분리균들은 형태학적 특성에 따라 무작위로 62개 집락을 선정하여 분리하였다. 순수 분리된 미생물들은 PCR 기법을 통해 16S rRNA 유전자의 염기서열을 분석한 후, 기존에 보고된 미생물 유전자 database와 비교함으로써 12속의 천일염 유래 호염균들의 존재를 확인하였다. 이번 연구 결과 천일염 생산 단계에서 분리된 halophilic bacteria은 Halobacillus, Halomonas, Bacillus, Idiomarina, Marinobacter, Pseudoalteromonas, Vibrio, Salinivibrio, Virgibacillus, Alteromonas, Staphylococcus 및 un-known 등이다.

Prognostic Significance of TP53 Mutations and Single Nucleotide Polymorphisms in Acute Myeloid Leukemia: A case Series and Literature Review

  • Zeichner, Simon Blechman;Alghamdi, Sarah;Elhammady, Gina;Poppiti, Robert John;Castellano-Sanchez, Amilcar
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권4호
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    • pp.1603-1609
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    • 2014
  • Background: The response to treatment and overall survival (OS) of patients with acute myeloid leukemia (AML) is variable, with a median ranging from 6 months to 11.5 years. TP53 is associated with old age, chemotherapy resistance, and worse OS. Using genetic sequencing, we set out to look at our own experience with AML, and hypothesized that both TP53 mutations and SNPs at codon 72 would mimic the literature by occurring in a minority of patients, and conferring a worse OS. Materials and Methods: We performed a pilot study of randomly selected, newly diagnosed AML patients at Mount Sinai Medical Center, diagnosed from 2005-2008 (n=10). TP53 PCR sequencing was performed using DNA from bone marrow smears. Analysis was accomplished using Mutation Surveyor software with confirmation of the variants using the COSMIC and dbSNP databases. Results: Fewer than half of the patients harbored TP53 mutations (40%). There was no significant difference in OS based on gender, AML history, risk-stratified karyotype, or TP53 mutation. There were possible trends toward improved survival among patients less than 60 (11 vs 4 months, p=0.09), Hispanics (8 vs 1 months, p=0.11), and those not harboring SNP P72R (8 vs 2 months, p=0.10). There was a significant improvement in survival among patients with better performance status (28 vs 4 months, p=0.01) and those who did not have a complex karyotype (8 vs 1 months, p=0.03). The most commonly observed TP53 mutation was a missense N310K (40%) and the most commonly observed SNP was P72R (100.0%). Conclusions: Our study confirms previous reports that poor PS and the presence of a complex karyotype are associated with a decreased OS. In our cohort, TP53 mutations were relatively common, occurring more frequently in male patients with an adverse karyotype. Although there was no significant difference in survival between TP53 mutated and un-mutated patients, there was a possible trend toward worse OS among patients with SNP P72R. Larger studies are needed to validate these findings.

Impact of Diet in Shaping Gut Microbiota Revealed by a Comparative Study in Infants During the First Six Months of Life

  • Fan, Wenguang;Huo, Guicheng;Li, Xiaomin;Yang, Lijie;Duan, Cuicui
    • Journal of Microbiology and Biotechnology
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    • 제24권2호
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    • pp.133-143
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    • 2014
  • The development of the gut is controlled and modulated by different interacting mechanisms, such as genetic endowment, intrinsic biological regulatory functions, environment influences and last but no least, the diet influence. In this work, we compared the fecal microbiota of breast-fed (BF), formula-fed (FF), and mixed-fed (MF) infants from Hebei Province, China. By using high-throughput 16S rDNA sequencing analyses, we found some differences in gut microbiota in the three groups. Firmicutes and Proteobacteria were the dominant bacteria at the phylum level in the three groups, where FF infants showed a significant depletion in Bacteroidetes (p < 0.001) and Actinobacteria (p < 0.05). Enterobacteriaceae was the dominant bacteria at the family level in the three groups, but FF infants showed higher Enterobacteriaceae enrichment than BF and MF infants (p < 0.05); the abundance of the Bifidobacteriaceae was only 8.16% in the feces of BF infants, but higher than in MF and FF infants (p < 0.05). The number of genera detected (abundance >0.01%) in BF, MF, and FF infants was only 15, 16, and 13, respectively. This study could provide more accurate and scientific data for the future study of infant intestinal flora.

Effects of husbandry systems and Chinese indigenous chicken strain on cecum microbial diversity

  • Dong, Xiuxue;Hu, Bing;Wan, Wenlong;Gong, Yanzhang;Feng, Yanping
    • Asian-Australasian Journal of Animal Sciences
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    • 제33권10호
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    • pp.1610-1616
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    • 2020
  • Objective: This study was to evaluate the effect of husbandry systems and strains on cecum microbial diversity of Jingyang chickens under the same dietary conditions. Methods: A total of 320 laying hens (body weight, 1.70±0.15 kg; 47 weeks old) were randomly allocated to one of the four treatments: i) Silver-feathered hens in enrichment cages (SEC) with an individual cage (70×60×75 cm), ii) Silver-feathered hens in free range (SFR) with the stocking density of 1.5 chickens per ten square meters, iii) Gold-feathered hens in enrichment cages (GEC), iv) Gold-feathered hens in free range (GFR). The experiment lasted 8 weeks and the cecum fecal samples were collected for 16S rDNA high throughput sequencing at the end of experiment. Results: i) The core microbiota was composed of Bacteroidetes (49% to 60%), Firmicutes (21% to 32%) and Proteobacteria (2% to 4%) at the phylum level. ii) The core bacteria were Bacteroides (26% to 31%), Rikenellaceae (9% to 16%), Parabacteroides (2% to 5%) and Lachnoclostridium (2% to 6%) at the genus level. iii) The indexes of operational taxonomic unit, Shannon, Simpson and observed species were all higher in SFR group than in SEC group while in GEC group than in GFR group, with SFR group showing the greatest diversity of cecum microorganisms among the four groups. iv) The clustering result was consistent with the strain classification, with a similar composition of cecum bacteria in the two strains of laying hens. Conclusion: The core microbiota were not altered by husbandry systems or strains. The free-range system increased the diversity of cecal microbes only for silver feathered hens. However, the cecum microbial composition was similar in two strain treatments under the same dietary conditions.

Purification and Characterization of a New Fibrinolytic Enzyme of Bacillus licheniformis KJ-31, Isolated from Korean Traditional Jeot-gal

  • Hwang, Kyung-Ju;Choi, Kyoung-Hwa;Kim, Myo-Jeong;Park, Cheon-Seok;Cha, Jae-Ho
    • Journal of Microbiology and Biotechnology
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    • 제17권9호
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    • pp.1469-1476
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    • 2007
  • Jeot-gal is a traditional Korean fermented seafood and has long been used for seasoning. We isolated 188 strains from shrimp, anchovy, and yellow corvina Jeot-gal, and screened sixteen strains that showed strong fibrinolytic activities on a fibrin plate. Among those strains, the strain that had the largest halo zone was chosen and identified as Bacillus licheniformis by using 16S rDNA sequencing and an API CHB kit. The fibrinolytic activity of Bacillus licheniformis was characterized and designated as bpKJ-31. The active component of bpKJ-31 was identified as a 37 kDa protein, designated bacillopeptidase F, by internal peptide mapping and N-terminal sequencing. The optimum activity of bpKJ-31 was shown at pH 9 and $40^{\circ}C$, with a chromogenic substrate for plasmin. It had high degrading activity for the $B{\beta}$-chain and $A{\alpha}$-chain of fibrin(ogen), and also acted on thrombin, but not skim milk and casein. The amidolytic activity of bpKJ-31 was inhibited by 1 mM phenylmethanesulfonyl fluoride, but 1 mM EDTA did not affect the enzyme activity, indicating that bpKJ-31 is an alkaline serine protease, like a plasmin. The bpKJ-31 showed approximately 14.3% higher fibrinolytic activity than the plasmin. These features of bpKJ-31 make it attractive as a health-promoting biomaterial.

Carrier frequency of SLC26A4 mutations causing inherited deafness in the Korean population

  • Kim, Hyogyeong;Lim, Hwan-Sub;Ryu, Jae-Song;Kim, Hyun-Chul;Lee, Sanghoo;Kim, Yun-Tae;Kim, Young-Jin;Lee, Kyoung-Ryul;Park, Hong-Joon;Han, Sung-Hee
    • Journal of Genetic Medicine
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    • 제11권2호
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    • pp.63-68
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    • 2014
  • Purpose: The mutation of the SLC26A4 gene is the second most common cause of congenital hearing loss after GJB2 mutations. It has been identified as a major cause of autosomal recessive nonsyndromic hearing loss associated with enlarged vestibular aqueduct and Pendred syndrome. Although most studies of SLC26A4 mutations have dealt with hearing-impaired patients, there are a few reports on the frequency of these mutations in the general population. The purpose of this study was to evaluate the prevalence of SLC26A4 mutations that cause inherited deafness in the general Korean population. Materials and Methods: We obtained blood samples from 144 Korean individuals with normal hearing. The samples were subjected to polymerase chain reaction to amplify the entire coding region of the SLC26A4 gene, followed by direct DNA sequencing. Results: Sequencing analysis of this gene identified 5 different variants (c.147C>G, c.225G>C, c.1723A>G, c.2168A>G, and c.2283A>G). The pathogenic mutation c.2168A>G (p.H723R) was identified in 1.39% (2/144) of the subjects with normal hearing. Conclusion: These data provide information about carrier frequency for SLC26A4 mutation-associated hearing loss and have important implications for genetic diagnostic testing for inherited deafness in the Korean population.

Lower Airway Microbiota and Lung Cancer

  • Sanchez-Hellin, Victoria;Galiana, Antonio;Zamora-Molina, Lucia;Soler-Sempere, Maria J.;Grau-Delgado, Justo;Barbera, Victor M.;Padilla-Navas, Isabel;Garcia-Pachon, Eduardo
    • 한국미생물·생명공학회지
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    • 제47권3호
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    • pp.441-448
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    • 2019
  • This study was aimed at identifying the lower airway microbiota in patients with lung cancer (LC) using protected brush sampling. We enrolled 37 patients undergoing diagnostic bronchoscopy for suspected LC, 26 with LC and 11 with benign diseases. Protected brush specimens were obtained from the contralateral lung and the side of the tumor; these specimens were analyzed by 16S rRNA-based-next-generation sequencing. The results indicated that the biodiversity was not different between groups, and there were no significant differences between the proportion of microorganisms in the tumor and in the contralateral side of patients with LC. In patients with LC, there was a higher abundance of several microorganisms including Capnocytophaga, Haemophilus, Enterococcus, and Streptococcus; whereas, in individuals without LC, Bacteroides, Lactobacillus, or Methylobacterium were more abundant. Malignancy could be determined with an accuracy of 70% by isolating Enterococcus, Capnocytophaga, or Actinomyces. Microbispora indicated benignity with a sensitivity of 55%, specificity of 88%, and accuracy of 78%. Lower airway microbiota in patients with LC is fairly similar in both the tumor and contralateral sites. Endobronchial microbiota is different in patients with and without LC, and these differences may have a potential clinical value as diagnostic or prognostic biomarkers.

Comparison of the oral microbial composition between healthy individuals and periodontitis patients in different oral sampling sites using 16S metagenome profiling

  • Kim, Yeon-Tae;Jeong, Jinuk;Mun, Seyoung;Yun, Kyeongeui;Han, Kyudong;Jeong, Seong-Nyum
    • Journal of Periodontal and Implant Science
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    • 제52권5호
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    • pp.394-410
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    • 2022
  • Purpose: The purpose of this study was to compare the microbial composition of 3 types of oral samples through 16S metagenomic sequencing to determine how to resolve some sampling issues that occur during the collection of sub-gingival plaque samples. Methods: In total, 20 subjects were recruited. In both the healthy and periodontitis groups, samples of saliva and supra-gingival plaque were collected. Additionally, in the periodontitis group, sub-gingival plaque samples were collected from the deepest periodontal pocket. After DNA extraction from each sample, polymerase chain reaction amplification was performed on the V3-V4 hypervariable region on the 16S rRNA gene, followed by metagenomic sequencing and a bioinformatics analysis. Results: When comparing the healthy and periodontitis groups in terms of alpha-diversity, the saliva samples demonstrated much more substantial differences in bacterial diversity than the supra-gingival plaque samples. Moreover, in a comparison between the samples in the case group, the diversity score of the saliva samples was higher than that of the supra-gingival plaque samples, and it was similar to that of the sub-gingival plaque samples. In the beta-diversity analysis, the sub-gingival plaque samples exhibited a clustering pattern similar to that of the periodontitis group. Bacterial relative abundance analysis at the species level indicated lower relative frequencies of bacteria in the healthy group than in the periodontitis group. A statistically significant difference in frequency was observed in the saliva samples for specific pathogenic species (Porphyromonas gingivalis, Treponema denticola, and Prevotella intermedia). The saliva samples exhibited a similar relative richness of bacterial communities to that of sub-gingival plaque samples. Conclusions: In this 16S oral microbiome study, we confirmed that saliva samples had a microbial composition that was more similar to that of sub-gingival plaque samples than to that of supra-gingival plaque samples within the periodontitis group.

Mutational Analysis of Korean Patients with Phenylketonuria

  • Koo, Soo Kyung;Lee, Kwang-Soo;Jung, Sung-Chul;Lee, Jong-Eun;Lee, Dong Hwan
    • 대한유전성대사질환학회지
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    • 제4권1호
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    • pp.5-12
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    • 2004
  • Purpose Phenylketonuria is an inborn error of metabolism, which is inherited as an autosomal recessive trait. PKU is resulting from deficiency of phenylalanine hydroxylase. PAH gene spans about 90 kb on chromosome 12q and comprises 13 exons. In order to define the genetic basis of PKU and the frequencies and distribution of PAH mutations in the Korean population, we analyzed PAH gene in independent 80 patients with PKU. Methods All 13 exons including exon-intron boundaries and 2 kb of 5' upstream region of the PAH gene were analyzed by PCR-direct sequencing methods. Results PAH gene analysis revealed 39 different mutations including 10 novel mutations. The novel mutations consisted of 9 missense mutations (P69S, G103S, N207D, T278S, P281A, L293M, G332V, S391I and A447P) and a novel splice site variant (IVS10-3C>G). R243Q, IVS4-1G>A, and E6-96A>G were the most relevant mutations and they accounted in the whole for 38% of the mutant alleles identified in this study. We also observed that. $BH_4$ responsibility was. associated with genotype of R241C, R53H and R408Q. Conc1ustion Our present study with 80 participants extends the previous results to more comprehensive understanding of PAH allele distribution and frequency in Koreans. Although Korean mutation profile of PAH is similar to those of the nearest oriental populations (Japanese, Chinese, and Taiwanese), several different characteristic features are revealed. The characterization of the genotype-phenotype relationship was also performed. Our data would be very useful information for diagnosis, genetic counseling and planning of dietary and therapeutic strategies in Korean PAH patients.

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돼지 Landrace 품종에서 Myostatin 유전자의 유전적 다형성과 성장형질과의 연관성 (Association Between MSTN Gene Polymorphism and Growth Traits in Landrace Pigs)

  • 조인철;최유림;고문석;김효선;이정규;전진태;한상현
    • Journal of Animal Science and Technology
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    • 제47권2호
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    • pp.159-166
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    • 2005
  • Porcine myostatin(MS1N) gene plays a key role in the differentiation of myoblast and muscle development. Genetic polymorphism was screened by single stranded conformation polymorphism(SSCP) analysis and subsequent DNA sequencing detected a nucleotide substitution(C2150T) in exon 3 of MSIN gene. Phenotypic association of the polymorphism was tested in a Landrace population and positive effects of the allele T for lean growth traits were found in the population. Even though it is not significant, the pigs have IT and TC genotypes were heavier for the body weight at birth and at twenty weeks of age than those containing genotype. Cc. However, the allele T was significantly associated with higher eye muscle area(P < 0.05). As a result of this study, we suggested that the allele T in exon 3 of MSTN gene comes a significant effect for increasing the eye muscle area without decreasing backfat thickness. This polymorphism did not change the amino acid but Taq I -RFLP matched to SSCP band patterns in exon 3 of MSTN gene, which will be an useful molecular marker for breeding of Landrace pigs.