• 제목/요약/키워드: Restriction Fragment Length

검색결과 539건 처리시간 0.03초

Use of Restriction Fragment Length Polymorphism Analysis to Differentiate Fungal Strains in Sunchang Meju

  • Jung, Jong-Hyun;Seo, Dong-Ho;Bhoo, Sung-Hee;Ha, Suk-Jin;Kim, Jong-Sang;Kim, Jeong-Hwan;Kwon, Dae-Young;Cha, Jae-Ho;Park, Cheon-Seok
    • Food Science and Biotechnology
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    • 제17권4호
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    • pp.888-891
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    • 2008
  • Twenty-three fungal strains were isolated from meju that had originated from the Sunchang province, the famous location for making fermented soybean foods in Korea. The restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rDNA (ITS-RFLP) was applied to differentiate the isolated fungal strains. First, the ITS region by polymerase chain reaction (PCR) with specific primers was amplified and then cleaved the products with different restriction enzymes. Cleavage of the amplified fragments with the restriction enzymes AluI, HaeIII, HhaI, and TaqI revealed extensive polymorphisms. The ITS-RFLP results highly correlated with ITS sequence analysis. All of the 23 fungal strains were classified into 5 groups by ITS-RFLP analysis. Aspergillus oryzae was the major fungal strain isolated from Sunchang meju (12 out of 23), while Aspergillus fumigatus was the next most frequently isolated strain (7 out of 23). In contrast, it was found that Fusarium asiaticum, Aspergillus sydowii, and Arthrinium sp. were the minor fungal strains in meju.

한우 혈액에서 PCR을 이용한 Mycobacterium avium ssp paratuberculosis의 검출 (Detection of Mycobacterium avium ssp paratuberculosis in Korean Cattle by the Polymerase Chain Reaction)

  • 김광현;곽길한;송희종;조정곤
    • 한국임상수의학회지
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    • 제27권1호
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    • pp.23-28
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    • 2010
  • Mycobacterium avium ssp paratuberculosis, intracellular bacteria that can cause chronic granulomatous enteritis in cattle, continues to pose significant economic losses and health problem with high prevalence. The purpose of this study is the polymerase chain reaction (PCR)-base strategy for early detection of M. avium ssp paratuberculosis in whole blood. Blood samples were collected from korean cattles in Jeonbuk, Korea. The 16 out of 88 serum samples were detected M. partuberculosis by ELISA. Then samples of infected 8 Korean cattles were amplified by PCR. The PCR amplified targets are 16s rDNA and heat shock protein 65kDa (hsp 65). The 16s rDNA provided a highly sensitive and specific tool for the direct detection of mycobacteria. In addition M. avium was confirmed characteristically by the hsp65. Finally there were sure to M. avium ssp paratuberculosis by IS900 PCR. The restriction fragment length polymorphism was identified by PCR amplifications and subsequence restriction enzyme digestions with Pst I of a hsp65. These results indicate that confirm M. avium with 16s rDNA, hsp65 and a restriction fragment length polymorphism in the hsp65 gene can be seem the other pattern. Therefore, these results can be used for clinical direct detections of M. avium ssp paratuberculosis in whole blood of Korean cattle and also to be used epidemiological researches.

Authentication of Salted-dried Fish Species Using Polymerase Chain Reaction-Single Strand Conformational Polymorphism and Restriction Analysis of Mitochondrial DNA

  • Kim, Joo-Shin;Chu, Kin Kan Astley;Kwan, Hoi Shan;Chung, Hau Yin
    • Fisheries and Aquatic Sciences
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    • 제11권3호
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    • pp.133-139
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    • 2008
  • Molecular techniques, including restriction fragment length polymorphism(RFLP) and polymerase chain reaction-single strand conformational polymorph isms(PCR-SSCP), were developed to identify salted, dried threadfin(Eleutheronema tetradactylum) and white herring(Ilisha elongata) fish. Using PCR with universal primers, conserved 367-bp fragments of the cytochrome b gene were amplified from fresh fish samples and sequenced. The sequences were then searched for specific restriction sites. The digestion of the PCR products with the endonucleases AvaI, FokI, MboII, and MspI generated RFLP, which was used to identify the commercial products. Similarly, the amplified PCR-SSCP products were developed and the products tested. Overall, similar patterns were found in the majority of the fresh and processed products. Based on the results, both RFLP and PCR-SSCP were useful in determining and validating the authenticity of the fish species used to prepare the commercial salted, dried products. A similar approach can be applied to other species.

Identification of Pork Contamination in Meatballs of Indonesia Local Market Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis

  • Erwanto, Yuny;Abidin, Mohammad Zainal;Muslim, Eko Yasin Prasetyo;Sugiyono, Sugiyono;Rohman, Abdul
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권10호
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    • pp.1487-1492
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    • 2014
  • This research applied and evaluated a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using cytochrome b gene to detect pork contamination in meatballs from local markets in Surabaya and Yogyakarta regions, Indonesia. To confirm the effectiveness and specificity of this fragment, thirty nine DNA samples from different meatball shops were isolated and amplified, and then the PCR amplicon was digested by BseDI restriction enzyme to detect the presence of pork in meatballs. BseDI restriction enzyme was able to cleave porcine cytochrome b gene into two fragments (131 bp and 228 bp). Testing the meatballs from the local market showed that nine of twenty meatball shops in Yogyakarta region were detected to have pork contamination, but there was no pork contamination in meatball shops in Surabaya region. In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity. Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked. The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

Identification of Quantitative Trait Loci Associated with Traits of Soybean for Sprout

  • Lee, Suk-Ha;Park, Keum-Yong;Lee, Hong-Suk;H. Roger Boerma
    • 한국작물학회지
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    • 제44권2호
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    • pp.166-170
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    • 1999
  • The identification of quantitative trait loci (QTL) has the potential to enhance the efficiency of im- proving food processing traits of soybean. In this study, 92 restriction fragment length polymorphism (RFLP) loci and two morphological markers (W$_1$ and T) were used to identify QTL associated with food processing traits of soybean for sprout in 83 F$_2$-derived lines from a cross of 'Pureun' x 'Jinpum 2'. The genetic map consisted of 76 loci which covered about 760 cM and converged into 20 linkage groups. Eighteen markers remained unlinked. Phenotypic data were collected for hypocotyl length, abnormal seedling rate, and sprout yield seven days after seed germination at 2$0^{\circ}C$. Based on the single-factor analysis of variance, eight independent markers were associated with hypocotyl length. Four of seven markers associated with abnormal seedling rate were identified as independent. Seven loci were associated with sprout yield. For three different traits, much of genetic variation was explained by the identified QTL in this population. Several RFLP markers in linkage group (LG) Bl were detected as being associated with three traits, providing a genetic explanation for the biological correlation of sprout yield with hypocotyl length (r=OA07***) and with abnormal seedling rate (r=-406***).

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유전자지문분석법(T-RFLP)을 이용한 하천 미생물의 다양성 평가 (Evaluation of Riverine Microbial Diversity using the Culture-Independent Genetic Fingerprinting Technique (T-RFLP))

  • 정주용;이경희
    • 한국물환경학회지
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    • 제24권2호
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    • pp.195-200
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    • 2008
  • To analyze the riverine microbial community structure, genetic fingerprints and ecological indexes such as species abundances, diversity, evenness, dominance of targeted rivers in Gyeonggi Province were acquired and evaluated using terminal restriction fragment length polymorphism (T-RFLP) technique. Genetic fingerprinting technique such as T-RFLP, which is able to show the microbial community clearly unlike traditional culture-dependent techniques, was thought to be useful to analyse the riverine microbial ecosystem under various factors. Riverine ecosystem evaluation using visible organisms would give biased results with time, targeted organism and researcher. But, T-RFLP, which can exclude the subjected biases such as culture condition and identification, would be an option to understand natural ecosystem by including the microorganisms that defy culture but perform important functions.

Nitrospira Community Composition in Nitrifying Reactors Operated with Two Different Dissolved Oxygen Levels

  • Park, Hee-Deung;Noguera, Daniel R.
    • Journal of Microbiology and Biotechnology
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    • 제18권8호
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    • pp.1470-1474
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    • 2008
  • Nitrospira is a dominant member of nitrite-oxidizing bacteria (NOB) in nitrifying bioreactors as well as in natural habitats. In this study, Nitrospira NOB were investigated in the two nitrifying reactors operated with high and low dissolved oxygen (DO) concentrations for a period of 300 days. Phylogenetic and terminal restriction fragment length polymorphism analyses based on 168 rRNA gene sequences revealed that the Nitrospira community compositions of the two reactors during the early period related to group 1 and half of the Nitrospira community composition shifted to group 2 in the high-DO reactor after day 179, although there was no significant change in the low-DO reactor. These results suggested that DO was an important factor affecting Nitrospira community compositions in the nitrifying reactors.