• 제목/요약/키워드: amplified fragment length polymorphism

검색결과 178건 처리시간 0.023초

Amplified Fragment Length Polymorphism Fingerprinting as a Tool to Study the Genetic Diversity of Staphylococcus aureus Isolated from Food Sources

  • Kim, Young-Sam;Kim, Jong-Bae
    • 대한의생명과학회지
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    • 제8권1호
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    • pp.39-46
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    • 2002
  • Amplified fragment length polymorphism (AFLP) is a recently developed PCR-based high resolution fingerprinting method that is able to generate complex banding patterns which can be used to delineate intraspecific genetic relationships among bacteria. In this study, we have modified and evaluated a PCR-based technique, amplified fragment length polymorphism (AFLP) analysis, for use in fingerprinting strains of Staphylococcus aureus. Single-enzyme amplified fragment length polymorphism (SE-AFLP) analysis was used to perform strain identification of Staphylococus aureus. By careful selection of AFLP primers, it was possible to obtain reproducible and sensitive identification to strain level. AFLP fingerprinting of 5 reference strains of Staphylococcus aureus and 65 strains of Staphylococcus aureus that were isolated from food sources of different area and diverse genomic types of Staphylococcus aureus were recognized. As a result of this study, we found that the AFLP patterns of Staphylococcus aureus isolated from Seoul, Taejeon and Gwang-Ju indicated the close relation with genetic similarity. The main purpose of this study was to find an alternative and reliable fingerprinting method to study the overall genetic diversity, using Staphylococcus aureus species as an example, and observed if the method can be successfully applied to all staphylococcal species.

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Fingerprinting of Listeria monocytogenes by Amplified Fragment Length Polymorphism Analysis

  • Jin, Hyun-Seok;Kim, Jong-Bae
    • 대한의생명과학회지
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    • 제8권1호
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    • pp.29-37
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    • 2002
  • Listeria monocytogenes poses an increasing health risk, which in part is due to increasing health risk, consumption of ready-to-eat food products and the introduction of increasing numbers of food products from regions with different dietary habits. L. monocytogenes can be present in meat, shellfish, vegetables, unpasteurised milk and soft cheese and poses a risk if food containing these products is stored at refrigeration temperature and is not properly heated before consumption, as L. monocytogenes is psychrophilic. Amplified-fragment length polymorphism (AFLP) analysis is the method of genotypic techinique in which adaptor oligonucleotides are ligated to restriction enzyme fragments and then used as target sites for primers in a PCR amplification. The amplified fragments are electrophoretically separated to give strain-specific band profiles. Single-enzyme approach that did not require costly equipment or reagents for the fingerprinting of strains of Listeria monocytogenes was developed. Single-enzyme amplified fragment length polymorphism (SE-AFLP) analysis was used to perform species and strain identification of Salmonella, Shigella, Yersinia and E. coli. By careful selection of AFLP primers, it was possible to obtain reproducible and sensitive identification to strain level. The AFLP patterns of L. monocytogenes are divided by the kinds of specimens in which were isolated. SE-AFLP fragments can be analyzed using standard gel electrophoresis, and can be easily scored by visual inspection, due to the low complexity of the fingerprint obtained by this method. These features make SE-AFLP suitable for use in either field or laboratory applications.

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Cleaved Amplified Polymorphic Sequence and Amplified Fragment Length Polymorphism Markers Linked to the Fertility Restorer Gene in Chili Pepper (Capsicum annuum L.)

  • Kim, Dong Sun;Kim, Dong Hwan;Yoo, Jae Hyoung;Kim, Byung-Dong
    • Molecules and Cells
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    • 제21권1호
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    • pp.135-140
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    • 2006
  • Cytoplasmic male sterility (CMS) in plants, which is due to failure to produce functional pollen, is a maternally inherited trait. Specific nuclear genes that suppress CMS, termed fertility restorer (Rf) genes, have been identified in several plants. In this study, Rfl-inked molecular markers in pepper (Capsicum annuum L.) were detected by bulked segregant analysis of eight amplified fragment length polymorphisms (AFLPs). Only AFRF8 was successfully converted to a cleaved amplified polymorphic sequence (CAPS) marker. This was named AFRF8CAPS and genotype determination using it agreed with that obtained with the original AFRF8. A linkage map with a total size of 54.1 cM was constructed with AFRF8CAPS and the seven AFLP markers using the Kosambi function. The AFRF8CAPS marker was shown to be closest to Rf with a genetic distance of 1.8 cM. These markers will be useful for fast and reliable detection of restorer lines during $F_1$ hybrid seed production and breeding programs in pepper.

Differentially Expressed Genes under Cold Acclimation in Physcomitrella patens

  • Sun, Ming-Ming;Li, Lin-Hui;Xie, Hua;Ma, Rong-Cai;He, Yi-Kun
    • BMB Reports
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    • 제40권6호
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    • pp.986-1001
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    • 2007
  • Cold acclimation improves freezing tolerance in plants. In higher plants, many advances have been made toward identifying the signaling and regulatory pathways that direct the low-temperature stress response; however, similar insights have not yet been gained for simple nonvascular plants, such as bryophytes. To elucidate the pathways that regulate cold acclimation in bryophytes, we used two PCR-based differential screening techniques, cDNA amplified fragment length polymorphism (cDNA-AFLP) and suppression subtractive hybridization (SSH), to isolate 510 ESTs that are differentially expressed during cold acclimation in Physcomitrella patens. We used realtime RT-PCR to further analyze expression of 29 of these transcripts during cold acclimation. Our results show that cold acclimation in the bryophyte Physcomitrella patens is not only largely similar to higher plants but also displays distinct differences, suggests significant alteration during the evolution of land plants.

Polymerase chain reaction - restriction fragment length polymorphism을 이용한 바이러스성 어류 질병 진단 (Diagnosis of viral fish diseases by polymerase chain reaction - restriction fragment length polymorphism)

  • 김명석;박신후;조미영;김진우;박명애
    • 한국어병학회지
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    • 제21권3호
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    • pp.181-188
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    • 2008
  • Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was used to detect and identify four fish viruses, fish iridovirus, viral hemorrhagic septicaemia virus (VHSV), viral nervous necrosis virus (VNNV), hirame rhabdovirus (HRV). Four viruses were detected by PCR with each specific primers. Identification of iridovirus was achieved by digesting the PCR amplified fragment with a restriction enzyme ApaⅠ. It was possible to distinguish positive from false positive PCR amplicons of VHSV by RFLP of PstⅠ or HindⅢ restriction enzymes. VNNV was identified using RFLP of BamHⅠrestriction enzyme and HRV was identified by XbaⅠ restriction enzyme. This approach can be used for more rapid, simple and specific diagnosis of fish viral diseases.

DNA Fingerprinting by Amplified Fragment Length Polymorphism Markers in Rainbow Trout(Oncorhynchus mykiss)

  • Yoon, Jong-Man;Park, Sang-Hoon
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2001년도 춘계 수산관련학회 공동학술대회발표요지집
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    • pp.559-560
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    • 2001
  • The objective of the present study was to analyze genetic variation and characteristics in rainbow trout(Oncorhynchus mykiss) using amplified fragment length polymorphism(AFLP) method as molecular genetic technique, to evaluate the usefulness of AFLP as genetic markers, and to compared the efficiency of agarose and polyacrylamide sequencing gels. The amplified products were performed by agarose and sequencing gel electrophoresis to detect AFLP band patterns, respectively. Using 9 primer combinations, total of 141 AFLP bands were produced, 108 bands(82.4%) of which were polymorphic in agarose gels. In sequencing gels, total of 288 bands were generated, and 220 bands (76.4%) were polymorphic. The level of bandsharing(BS) ranged from 0.18 to 0.32 for the 9 primer combinations tested, with a mean of 0.24. Consequently, AFLP markers of these rainbow trout could be used as genetic information such as species identification, genetic relationship or analysis of genome structure, and selection aids for genetic improvement of economically importment traits in fish species.

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Identification and characterization of novel single nucleotide polymorphism markers for fat deposition in muscle tissue of pigs using amplified fragment length polymorphism

  • Supakankul, Pantaporn;Kumchoo, Tanavadee;Mekchay, Supamit
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권3호
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    • pp.338-346
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    • 2017
  • Objective: This study was conducted to identify and evaluate the effective single nucleotide polymorphism (SNP) markers for fat deposition in the longissimus dorsi muscles of pigs using the amplified fragment length polymorphism (AFLP) approach. Methods: Sixty-four selective primer combinations were used to identify the AFLP markers in the 20 highest- and 20 lowest-intramuscular fat (IMF) content phenotypes. Five AFLP fragments were converted into simple codominant SNP markers. These SNP markers were tested in terms of their association with IMF content and fatty acid (FA) composition traits in 620 commercially crossbred pigs. Results: The SSC7 g.4937240C>G marker showed an association with IMF content (p<0.05). The SSC9 g.5496647_5496662insdel marker showed a significant association with IMF content and arachidonic levels (p<0.05). The SSC10 g.71225134G>A marker revealed an association with palmitoleic and ${\omega}9$ FA levels (p<0.05), while the SSC17 g.61976696G>T marker showed a significant association with IMF content and FA levels of palmitoleic, eicosenoic, arachidonic, monounsaturated fatty acids, and ${\omega}9$ FA levels. However, no significant association of SSC8 g.47338181G>A was observed with any IMF and FA levels in this study. Conclusion: Four SNP markers (SSC7 g.4937240C>G, SSC9 g.5496647_5496662insdel, SSC10 g.71225134G>A, and SSC17 g.61976696G>T) were found to be associated with IMF and/or FA content traits in commercially crossbred pigs. These findings provide evidence of the novel SNP markers as being potentially useful for selecting pigs with the desirable IMF content and FA composition.

Population Genetic Structure of Carassius auratus (Pisces: Cypriniformes) in South Korea Inferred from AFLP Markers: Discordance with Mitochondrial Genetic Structure

  • Jung, Jongwoo
    • Animal Systematics, Evolution and Diversity
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    • 제29권1호
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    • pp.18-22
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    • 2013
  • A recent study on the mitochondrial genetic variation of the Carassius auratus population in South Korea suggested that there are 3 distinct mitochondrial lineages in the country, and that they are geographically separated between westward rivers and southward rivers, respectively. In this study, the population genetic structure of amplified fragment length polymorphism (AFLP) of Carassius auratus was investigated. The results of analysis of molecular variance (AMOVA) supported the geographic distinction between westward and southward river populations, but only 3.66% of total genetic variance lies among these populations. The panmicticity of the AFLP genetic variation is backed up by the results of the neighbor-joining dendrogram drawn from a linearized pairwise $F_{ST}$ matrix and Bayesian clustering analysis. The discordance of genetic structure between mitochondrial and AFLP genetic variation may come from difference in effective population size between these markers and/or gene flow between westward and southward river populations through river capture events.