• Title/Summary/Keyword: t-RFLP

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Identification of Beef Breed using DNA Marker of Coat Color Genes (모색 발현 유전자의 DNA Marker를 이용한 쇠고기 품종 판별)

  • Chung Eui-Ryong;Chung Ku-Young
    • Food Science of Animal Resources
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    • v.24 no.4
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    • pp.355-360
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    • 2004
  • In Korean beef market, one of the major problems is mislabeling or fraudulent distribution of Holstein dairy meat or imported beef as domestic Hanwoo meat. Therefore, there has been a great need for a development of technology to identify beef breeds in meat and meat products. This study was carried out to develop the accurate and reliable method for the identification of beef breed using PCR-RFLP marker of MC1R, MGF and TYRPl genes affecting coat colors in cattle. A single base substitution (G\longrightarrowT transition) at the codon for amino acid position 104 of MC1R gene was identified between Hanwoo and Holstein and Angus breeds. The change at this position creates Msp I restriction site in Holstein and Angus, but not in Hanwoo. When the DNA amplified products (537 bp) was digested with Msp I, Hanwoo meat showed a single band of 537bp, while two fragments of 329bp and 208 bp were observed in Holstein meat and Angus breed, respectively. Thus, breed-specific RFLP marker in the MC1R gene can be used to distinguish between Hanwoo meat and Holstein and Angus meats. In the RFLP genotype of MGF gene, the frequency of r/r type was 75% in Manwoo, whereas the frequency of R/R was 80% in Hereford breed. Holstein and Angus breeds showed 100% for R/r type. Therefore, Hanwoo meat showed significant difference in the MGF genotype frequencies compared with those of Holstein meat and imported beef cattle breeds. However, TYRP1 gene showed the same genotype in all breeds examined. Thus, this TYRP1 gene can not be used as a molecular marker for breed identification. As a consequence, we suggest that RFLP markers of the MC1R and MGF coat color genes could be used as DNA marker for identification of Hanwoo meat from Holstein and imported meats.

Phylogenetic Study of Trichaptum Species Based on the RFLP Analysis of Mitochondrial DNA

  • Kim, Mi-Sun;Jung, Hack-Sung
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.215-219
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    • 1996
  • Eight strains of Trichaptun (Polyporaceae), two strains from each species of T. abietinum, T. biforme, T. fusco-violaceum, and T. laricinum were examined to see their phylogenetic relationship by digesting mitochondrial DNAs with EcoRV, Hind III, XbaI, and PstI, and then analyzing fragmentation patterns with the methods of Nei and Li. T. abietinum, T. biforme, and T. laricinum developed an independent phylogenetic lineage, respectively, but T. fusco-violaceum FP-133997-sp showed a close relationship with two strains of T. bioforme, and T. fusco-violaceum HHB-4016-sp barely grouped with those of T. laricinum. Based on the results of the RFLP analysis of mtDNA, it is concluded that T. fusco-violaceum is under way to differentiation into two different subgroups.

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Comparative Analysis of Cyanobacterial Communities from Polluted Reservoirs in Korea

  • Kim, Jin-Book;Moon, Mi-Sook;Lee, Dong-Hun;Lee, Sung-Taik;Bazzicalupo, Marco;Kim, Chi-Kyung
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.181-187
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    • 2004
  • Cyanobacteria are the dominant phototrophic bacteria in water environments. Here, the diversity of cyanobacteria in seven Korean reservoir waters where different levels of algal blooms were observed during the summer of 2002, was examined by T-RFLP analysis. The number of T-RF bands in the HaIII T-RFLP profiles analyzed from those water samples ranged from 20 to 44. Of these, cyanobacteria accounted for 6.1 to 27.2% of the total bacteria. The water samples could be clustered into 2 groups according to the Dice coefficient of the T -RF profiles. The eutrophic Dunpo and oligotrophic Chungju reservoirs were selected, and several representative clones from both reservoir waters analyzed for the nucleotide sequences of their 16S rDNA. The major clones were found to belong to the Microcystis and Anabaena species in the waters from the Dunpo and Chungju reservoirs, respectively, which was in agreement with the T-RFLP result. That is, the Microcystis and Anabaena species were dominant in the eutrophic and polluted Dunpo and oligotrophic Chungju reservoir waters, respectively. These results indicated that there is a correlation between prevalence of cyanobacterial species and levels of pollution in reservoir waters.

Monitoring of Microorganisms Added into Oil-Contaminated Microenvironments by Terminal-Restriction Fragment Length Polymorphism Analysis

  • JUNG SEONG-YOUNG;LEE JUNG-HYUN;CHAI YOUNG-GYU;KIM SANG-JIN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1170-1177
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    • 2005
  • Terminal-restriction fragment length polymorphism (T-RFLP) analysis was used to monitor inoculated oil-degrading microorganisms during bioremedial treatability tests. A pair of universal primers, fluorescently labeled 521F and 1392R, was employed to amplify small subunit rDNA in order to simultaneously detect two bacterial strains, Corynebacterium sp. IC10 and Sphingomonas sp. KH3-2, and a yeast strain, Yarrowia lipolytica 180. Digestion of the 5'-end fluorescence/labeled PCR products with HhaI produced specific terminal-restriction fragments (T-RFs) of 185 and 442 bases, corresponding to Corynebacterium sp. IC10 and Y. lipolytica 180, respectively. The enzyme NruI produced a specific T-RF of 338 bases for Sphingomonas sp. KH3-2. The detection limit for oildegrading microorganisms that were inoculated into natural environments was determined to be $0.01\%$ of the total microbial count, regardless of the background environment. When three oil-degrading microorganisms were released into oil-contaminated sand microenvironments, strains IC10 and 180 survived for 35 days after inoculation, whereas strain KH3-2 was detected at 8 days, but not at 35 days. This result implies that T-RFLP could be a useful tool for monitoring the survival and relative abundance of specific microbial strains inoculated into contaminated environments.

Bacterial Community Structure and Diversity Using 16S rDNA Analysis in the Intertidal Sediment of Ganghwa Island (16S rDNA 분석을 이용한 강화도 장화리 갯벌 퇴적물 내 미생물 군집구조 및 다양성)

  • Cho Hye Youn;Lee Jung-Hyun;Hyun Jung-Ho
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.189-198
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    • 2004
  • T-RFLP analysis and clone sequencing analysis based on bacterial 16S rDNA were conducted to assess bacterial community structure and diversity in two layers (0-1cm, 6-7cm depth) of the sediment from Janghwari intertidal flat in Ganghwa Island. The results of T-RFLP (terminal-restriction fragment length polymorphism) analysis using restriction enzyme HhaI showed that the T-RFs of various size ($60{\pm}2$) bp-($667{\pm}2$) bp) appeared evenly at the surface sediments but two T-RFs with 60(${\pm}2$)bp and 93 (${\pm}2$)bp predominated at 6-7cm depth. Analysis of partial sequences for 172 clones revealed that 98% of the clones were not matched with the sequences of cultured bacteria strains in the GenBank (${\geq}similarity$ 98%), and approximately 86% of them were classified as different phylotypes. Most clones belonged to $\alpha$-, $\gamma$-, and $\delta$-Proteobacteria, Acidobacteria/Holophaga and green nonsulfur bacteria group. Proteobacteria group occupied the highest proportion in both layers (69% at 0-1cm depth and 46% at 6-7cm depth). $\gamma$-Proteobacteria and $\delta$-Proteobacteria that are associated with oxidation and reduction of sulfur compounds were appeared to be dominant, and comprised 21.5% and 15.7% of total clones, respectively. Overall results indicated that extremely diverse bacterial groups were inhabiting in the sediment of Ganghwa intertidal flat, and bacterial communities associated with the behaviour of sulfur seemed to playa significant role in the biogeochemical environment in this anoxic sediment.

PAHs로 오염된 광양만 퇴적토의 미생물 군집분석

  • 권개경;정성영;이정현;바실리 스베타체프;김상진
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2002.11a
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    • pp.51-58
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    • 2002
  • 연안에 가까운 광양만 퇴적토를 대상으로 하여 PAHs 오염도와 말단제한절편 다형성 분석방법(T-RFLP method)을 이용하여 미생물 군집구조를 조사하였다. 조사된 10개 정점의 PAHs오염도는 80$\pm$20~5,690$\pm$580 (평균 1,499$\pm$1,716) ng/g dry wt.였으며 폐수종말처리장이 위치하는 하천 입구에 위치한 정점 (Wl)과 광양제철소 우수배출구 입구에 위치한 정점에서 상대적으로 높은 농도를 보였다. T-RFLP방법으로 조사한 미생물 군집은 10개 정점이 4개의 그룹으로 구분되었으며 정점 Wl에서 다른 정점과 다른 독특한 군집구조를 형성하였다. PAHs 농후배양시료에서의 미생물군집구조 변화와 비교해 볼 때 조사대상지역 퇴적토의 미생물 군집구조는 PAHs의 오염에 부분적으로는 영향을 받지만 입도, 유기물 등과 같은 환경요인이 군집구조를 결정하는 주된 요인인 것으로 생각된다.

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Two-year field monitoring shows little evidence that transgenic potato containing ABF3 significantly alters its rhizosphere microbial community structure

  • Nam, Ki Jung;Kim, Hyo-Jeong;Nam, Kyong-Hee;Pack, In Soon;Kim, Soo Young;Kim, Chang-Gi
    • Journal of Ecology and Environment
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    • v.41 no.3
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    • pp.99-106
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    • 2017
  • Background: Plants over-expressing Arabidopsis ABF3 (abscisic acid-responsive element-binding factor 3) have enhanced tolerance to various environmental stresses, especially drought. Using terminal restriction fragment length polymorphism (T-RFLP) analysis, we compared the rhizosphere-associated structures of microbial communities for transgenic potato containing this gene and conventional "Jopoong" plants. Results: During a 2-year field experiment, fungal richness, evenness, and diversity varied by year, increasing in 2010 when a moderate water deficit occurred. By contrast, the bacterial richness decreased in 2010 while evenness and diversity were similar in both years. No significant difference was observed in any indices for either sampling time or plant line. Although the composition of the microbial communities (defined as T-RF profiles) changed according to year and sampling time, differences were not significant between the transgenic and control plants. Conclusions: The results in this study suggest that the insertion of ABF3 into potato has no detectable (by current T-RFLP technique) effects on rhizosphere communities, and that any possible influences, if any, can be masked by seasonal or yearly variations.

Relationship between the Polymorphisms of 5' Regulation Region of Prolactin Gene and Milk Traits in Chinese Holstein Dairy Cows

  • Li, J.T.;Wang, A.H.;Chen, P.;Li, H.B.;Zhang, C.S.;Du, L.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.4
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    • pp.459-462
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    • 2006
  • Prolactin (PRL) plays an important role in promoting mammalian mammary gland development, and milk production during lactation. Therefore the PRL gene was chosen as a candidate gene for milk traits in Holstein dairy cows. PCR-SSCP and PCR-RFLP were used to analyze genetic variations in the 5' regulation region of the PRL gene. In this part of the gene, two new polymorphic sites were detected in the Chinese Holstein dairy cows. One was a XbaI-RFLP locus, and the other was an SSCP locus. Statistical analysis showed that the XbaI-RFLP locus and the SSCP locus had a significant positive effect on milk traits.