• Title/Summary/Keyword: T-RFLP

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Evaluation of the Colonization of Lactobacillus plantarum in Mouse Gut by Terminal Restriction Fragment Length Polymorphism Analysis (Terminal Restriction Fragment Length Polymorphism 분석을 이용한 Lactobacillus plantarum의 생쥐 장관 정착 평가)

  • Jung, Gwangsick;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.389-395
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    • 2012
  • T-RFLP (terminal restriction fragment length polymorphism) analysis, one of the most highly adopted culture-independent microbial community analysis methods, was applied to evaluate the colonization of probiotics in experimental animal gut. Lactic acid bacteria that exhibited cinnamoyl esterase activity were isolated from Korean fermented vegetables and identified by 16S ribosomal RNA sequence analysis. Lactobacillus plantarum KK3, which demonstrated high chlorogenic acid hydrolysis by cinnamoyl esterase activity, and acid/bile salt resistances, was cultured, freeze-dried, and fed to mice and the microbiota in their feces were monitored by T-RFLP analysis. The T-RF of L. plantarum was detected in the feces of mice after the start of administration and lasted at least 31 days after the initial 7 day feeding. T-RFLP analysis was considered a useful tool to evaluate the gut colonization of probiotic L. plantarum. In order to prove that L. plantarum was from viable cells, we reisolated L. plantarum in the feces using cinnamoyl esterase activity media as the screening step. The colonization of L. plantarum KK3 in the mouse gut was confirmed by this research.

Profile Analysis of Bacteria in Human Hands Using the Terminal Restriction Fragment Length Polymorphism (T-RFLP) Analysis (제한효소 절편길이 다형성(T-RFLP) 분석기법을 이용한 손에 서식하는 세균의 군집조성 분석)

  • Park, Jisun;Kim, Seung Bum
    • Journal of Science Criminal Investigation
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    • v.11 no.4
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    • pp.276-282
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    • 2017
  • As evidence that can be obtained at the crime scene, the importance of micro-evidences has been recognized in recent years with the development modern molecular-level analytical techniques. These micro-evidences include substances useful for personal identification such as DNA, but it is difficult to collect only the evidences showing individual characteristics every time at the crime scene. Therefore, development of new research approaches for the discovery and application of micro-evidence candidates is in increasing demand. For this purpose, skin microbial communities of bacteria inhabiting the palms of 16 people were collected and terminal-restriction enzyme fragment length polymorphism (T-RFLP) analysis was carried out to examine the potential for the application in personal identification. As a result, 16 different electropherograms were obtained, and various taxa including Staphylococcus and Bacillus were shown to produce different T-RF profiles among individuals. These results were analyzed with the factors affecting the microbiota such as sex and working environment of individuals.

Methane Production and T-RFLP Patterns of Methanogenic Bacteria Dependent on Agricultural Methods (농법에 따른 메탄생성과 메탄생성 세균의 T-RFLP 패턴)

  • Kim, Hun-Soo;Cho, Ju-Sik;Park, Kyeong-Ryang
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.17-25
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    • 2009
  • We studied soil components, methane production, the number of methanogens, and T-RFLP patterns dependent on agricultural methods with the change of seasons. There is no regular increase or decrease tendency of the most soil components followed by sampling period. And the water content in soil was higher in October than May. Also a lot of methanogens existed in soil, and acetotrophs were relatively of smaller number than hydogenotrophs and formate utilizing methanogens using MPN (most probable number) enumeration. In the experiment using the formate, it was used from the first week, and only a minute amount was detecte after four weeks. However in the acetate, it was increased until the third week, and after that was consumed. And there was higher methane production for all soil samples which administered with the hydrogen spike. The activity of methanogens was higher in the organic and low-agrichemical agricultural method samples, and the organic agricultural method had high methanogen activity among the other samples. A result of T-RFLP pattern of mcrA gene digested with Sau96I, methanogen community have a little relation with agricultural methods and seasons. This results also agreed to no critical difference the soil components dependent on agricultural methods, but some analytical data have a positive relationship with a agricultural methods. Therefor we could concluded that the comparison study of community for soil bacteria sufficiently could be useful for the microbiological indicator.

Effects of Polycyclic Aromatic Hydrocarbons Contamination on the Community Structure of Microorganisms in Gwangyang bay Sediments (다환방향족 탄화수소가 광양만 퇴적토의 미생물 군집구조에 미치는 영향)

  • 권개경;정성영;이정현;현정호;김상진
    • Korean Journal of Environmental Biology
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    • v.22
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    • pp.38-46
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    • 2004
  • Effect of polycyclic aromatic hydrocarbons (PAHs) on the community structure of indigenous microorganisms in Gwangyang Bay sediments was investigated in Mar. & Aug.,2000. Microbial community structure was analyzed using 5'-terminal restriction fragment length polymorphism (T-RFLP) method. Microbial community structure based on T-RFLP method revealed that community differentiated by sampling period except station 1 located near the stream discharge site from Yeosu Industrial Complex. Even, microbial diversity was higher at stations showed relatively high concentrations of PAHs. The microbial community structure was severely changed during the enrichment culture with 1,000 ppm of PAHs mixture. It was also different between cultivated at 8$^{\circ}C$ and 30$^{\circ}C$. The results implied that temperature, poyosity, organic content and etc were more responsible than PAHs on the microbial community structure.

Comparison of community structure of sulfate reducing bacteria in rice paddy and dry farming soils (논과 밭 토양의 황산염 환원세균 군집 구조 비교)

  • Lee, Jung Bae;Park, Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.21-30
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    • 2015
  • The goal of this study was to identify relationships between the composition of sulfate reducing bacterial assemblages and terminal restriction fragment length polymorphism (T-RFLP) patterns in rice paddy and dry farming soils. Samples of organic farming soils, conventional farming soils, and dry field farming soils were collected in August and November. Analyses of the soil chemical composition revealed similar total nitrogen, total carbon and total inorganic phosphorus levels; however, the moisture content and total carbon were higher than in the other soils in both August and November, respectively. Sulfate reducing bacteria utilizing lactic acid were more widely distributed than those that used acetic acid, and the number of sulfate reducing bacteria in organic farming soil was most abundant. Phylogenetic analysis based on 181 clones revealed that most showed low similarity with cultured sulfate reducing bacteria, but more than 90% similarity with an uncultured sulfate reducing bacteria isolated from the environment. T-RFLP analysis revealed that fragments of 91, 357, 395, and 474 bp were most common, and the community structure of sulfate reducing bacteria changed seasonally.

Association Study between Vitamin D Receptor Gene Polymorphism and Adult Periodontitis in Korea

  • Kang, Byung-Yong;Ha, Nam-Joo
    • Animal cells and systems
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    • v.7 no.2
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    • pp.145-149
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    • 2003
  • Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RFLP) in the vitamin D receptor gene in nan periodontically healthy controls and 28 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene was detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There was no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.

Association of Sequence Variations in DGAT 1 Gene with Economic Traits in Hanwoo (Korea Cattle)

  • Kong, H.S.;Oh, J.D.;Lee, J.H.;Yoon, D.H.;Choi, Y.H.;Cho, B.W.;Lee, H.K.;Jeon, G.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.817-820
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    • 2007
  • The effects of diacylglycerol O-acyltransferase (DGAT1) candidate gene polymorphism on the economic traits of Hanwoo were studied. Through sequencing analysis, two polymorphism sites at K232A and T11993C were established and were analyzed by PCR-RFLP. The PCR-RFLP analysis for K232A showed that the frequencies of alleles K and A were 0.75 and 0.25, respectively, and the frequencies of genotypes for K/K, K/A and A/A were estimated as 0.509, 0.491 and 0, respectively. In the PCR-RFLP analysis for T11993C, we found allele frequencies of 0.773 and 0.227 for T and A, respectively, and 0.546, 0.454 and 0 for the T/T, T/C and C/C genotype frequencies, respectively. No significant effects on economic traits in Hanwoo were found in the separate analysis of K232A and T11993C polymorphisms, but the interaction between K232A and T11993C showed a significant effect (p<0.005) on marbling score. The DGAT1 candidate gene was found to have a significant effect not only on milk yield and component traits but also on the metabolism of intramuscular fat.

Monitoring of Horizontal Gene Transfer from Agricultural Microorganisms to Soil Bacteria and Analysis of Microbial Community in Soils

  • Kim, Sung-Eun;Moon, Jae-Sun;Choi, Won-Sik;Lee, Sang-Han;Kim, Sung-Uk
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.563-566
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    • 2012
  • To investigate the possibility of horizontal gene transfer between agricultural microorganisms and soil microorganisms in the environment, Bacillus subtilis KB producing iturin and the PGPR recombinant strain Pseudomonas fluorescens MX1 were used as model microorganisms. The soil samples of cucumber or tomato plants cultivated in pots and the greenhouse for a six month period were investigated by PCR, real-time PCR, Southern hybridization, and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Our data from Southern blotting and T-RFLP patterns suggest that the model bacteria do not give significant impacts on the other bacteria in the pots and greenhouse during cultivation.

Genetic Polymorphisms of t-PA and PAI-1 Genes in the Korean Population

  • Kang, Byung-Yong;Lee, Kang-Oh
    • Animal cells and systems
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    • v.7 no.3
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    • pp.249-253
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    • 2003
  • Abnormalities in fibrinolysis system is associated with risk of hypertension. In this report, the Alu repeat insertion/deletion (I/D) polymorphism of tissue plasminogen activator (t-PA) and the Hind III RFLP of plasminogen activator inhibitor-1 (PAI-1) genes were investigated in 115 normotensives and 83 patients with hypertension, and their association with anthropometrical data and plasma biochemical parameters were analyzed. There were no significant differences in the gene frequencies of the two candidate genes between normotensives and hypertensives, respectively. Our results indicate lack of associations between the two polymorph isms in t-PA and PAI-1 genes and risk of hypertension in the population under study. However, the Hind III RFLP of PAI-1 gene was significantly associated with plasma glucose level, suggesting its role in glucose metabolism. It needs to be tested whether this RFLP of PAI-1 gene is associated with insulin resistance syndrome or non-insulin dependent diabetes mellitus (NIDDM) in the Korean population.