• Title/Summary/Keyword: DNA fingerprinting

Search Result 149, Processing Time 0.026 seconds

Retrotransposon Microsatellite Amplified Polymorphism Strain Fingerprinting Markers Applicable to Various Mushroom Species

  • Le, Quy Vang;Won, Hyo-Kyung;Lee, Tae-Soo;Lee, Chang-Yun;Lee, Hyun-Sook;Ro, Hyeon-Su
    • Mycobiology
    • /
    • v.36 no.3
    • /
    • pp.161-166
    • /
    • 2008
  • The retrotransposon marY1 is a gypsy family retroelement, which is detected ubiquitously within the fungal taxonomic groups in which mushrooms are included. To utilize marY1 as a molecular marker for the DNA fingerprinting of mushrooms, oligonucleotides marY1-LTR-L and marY1-LTR-R were designed on the basis of highly conserved regions from the multiple sequence alignment of 30 marY1 sequences retrieved from a nucleotide sequence database. In accordance with $\underline{Re}trotransposon$ $\underline{M}icrosatellite$ $\underline{A}mplified$ $\underline{P}olymorphism$ (REMAP) fingerprinting methodology, the two oligonucleotides were utilized together with the short sequence repeat primers UBC807 and UBC818 for polymerase chain reaction using templates from different mushroom genomic DNAs. Among the tested oligonucleotides, the marY1-LTR-L and UBC807 primer set yielded the greatest amount of abundance and variation in terms of DNA band numbers and patterns. This method was successfully applied to 10 mushroom species, and the primer set successfully discriminated between different commercial mushroom cultivars of the same strains of 14 Pleurotus ostreatus and 16 P. eryngii. REMAP reproducibility was superior to other popular DNA fingerprinting methodologies including the random amplified polymorphic DNA method.

Toxin Gene Typing, DNA Fingerprinting, and Antibiogram of Clostridium perfringens Isolated from Livestock Products

  • Lee, Seung-Bae;Choi, Suk-Ho
    • Food Science of Animal Resources
    • /
    • v.26 no.3
    • /
    • pp.394-401
    • /
    • 2006
  • Forty Clostridium perfringens isolates were obtained from twelve animal products, following the examination of eighty six beef, pork, broiler chicken and salami meat products, and eleven milk powder products. There were 21 isolates from salami stored at $25^{\circ}C$, 3 isolates from pork, 4 isolates from beef, 9 isolates from broiler chicken, and 3 isolates from milk powder. Only the cpa gene encoding a toxin among the 5 toxin genes tested (cpa, cpb, etx, iap, and cpe) was detected in all forty isolates, suggesting contamination with C. perfringens type A. DNA fingerprinting analysis using PCR of the tRNA intergenic spacer (tDNA-PCR) and the 16S-23S internal transcribed spacer (ITS-PCR), and randomly amplified polymorphic DNA (RAPD) analysis were attempted to differentiate the isolates. RAPD analysis was the most discriminating method among the three PCR analyses. Isolates from the same products tended to show similar RAPD patterns. Antimicrobial susceptibility tests showed that some isolates from broiler chickens had the same antibiogram with multiple resistance to streptomycin, colistin, and ciprofloxacin. Antibiograms were similar between isolates from the same livestock products, but differed considerably between the products.

Genomic Fingerprinting of Staphylococcus aureus Isolated from Bovine Mastitis Milk by PCR (젖소 유방염 유래 Staphylococcus aureus의 PCR을 이용한 Genomic Fingerprinting)

  • Kim, Doo;Kwon, Soon-Tak;An, So-Jeo
    • Journal of Veterinary Clinics
    • /
    • v.16 no.1
    • /
    • pp.19-25
    • /
    • 1999
  • A total of 137 strains of Staphylococcus aureus were isolated from dairy cow's milk with subclinical mastitis from 33 herds in 5 provinces and 36 strains of S aureus from clinical mastitis from 4 herds where the mastitis were severe problem. Arbitrary primed polymerase chain reactions with 10 bp oligonucleotide primer were performed and the PCR products were analysed with image analyzer, The S aureus strains were genotyped into 20 distinct DNA fingerprinting profiles. The size of PCR products ranged from 163 to 2,479 bp and PCR products of 506, 770, 784 and 2,479 bp were the most prevailing bands. Genotype 3 was founded in all 5 provinces. The various genotypes were identified in newly founded dairy herds, however, only one or two genotypes were identified in the closed herds. In clinical mastitis, only a limited number of different S aureus genotype was founded in each of the herds in comparision with subclinical mastitis. The results demonstrated that PCR-based DNA fingerprinting analysis of S aureus strain can be used to study epidemiology of mastitis, in addition, common genotype in geographic region can be useful for the development of an effective S aureus bacterin.

  • PDF

Optimization of the Concentrations of ERIC-PCR Components to Simultaneously Differentiate Five Foodborne Pathogenic Bacterial Genera (식중독세균 5속의 동시 동정을 위한 ERIC-PCR 반응성분 농도의 최적화)

  • Seo, Hyun-Ah;Park, Sung-Hee;Kim, Keun-Sung
    • Journal of Food Hygiene and Safety
    • /
    • v.18 no.4
    • /
    • pp.229-236
    • /
    • 2003
  • The five different foodborne pathogenic bacterial genera of Escherichi, Salmonella, Shigella, Vibrio and Listeria are important sources of foodpoison. However, the method was not developed to simultaneously differentiate these five bacteria at molecular level. The optimized concentrations of the four major PCR cocktail components of $MgCl_2$, dNTPs, primers and template DNA were determined when ERIC (enterobacterial repetitive intergenic consensus)-PCR reactions were carried out to differentiate the five differnet foodborne pathogenic bacteria. The optimized concentration of $MgCl_2$ was determined to be 2 mM in order to obtain a consistent fingerprinitng pattern. The similar fingerprinting pattern was obtained when ERIC primers and dNTPs were added up to the concentrations of 2 ${\mu}M$ and 200 ${\mu}M$, respectively. As for template DNA, the numbers of PCR fragments were not affected, but their intensities were increased as the concentrations of the DNA were increased.

Comparison of Polymorphisms of Salmonella pullorum and Salmonella gallinarum Using DNA Amplified Fingerprinting (DNA Amplified Fingerprinting 기법을 이용한 Salmonella pullorum과 Salmonella gallinarum의 다형성 비교 분석)

  • Kim, Yeon-Soo;Kim, Sang-Kyun;Song, Won-Chul;Hwang, Eui-Kyung
    • Korean Journal of Veterinary Research
    • /
    • v.41 no.3
    • /
    • pp.357-365
    • /
    • 2001
  • This study was performed to detect the Salmonella genus-specific DNA marker for comparing of polymophisms between S pullorum and S gallinarum by using PCR amplified techniques. A total of ten primers were used to detect DNA polymorphisms from S pullorum and S gallinarum. The number of DAF bands detected per each primer varied from 26 to 45, with an average of 32.7 using 10 primers. A total of 327 DAF bands were generated and among them 123 bands were polymorphic(37.6%). These DNA amplified fingerprinting(DAF) specific bands for S pullorum and S gallinarum were observed from all primers. For S pullorum, GEN 60-04, GEN 70-04 and GEN 70-03 primers showed a high level of polymorphism with 0.79, 0.70 and 0.57, respectively. But GEN 60-05 primer did not show a level of polymorphism. For S gallinarum, GEN 70-03, 60-04, 60-07, 70-05 and 70-04 primers showed a higher a low level of polymorphism from 0.16 to 0.28. Each five strains of S pullorum and S gallinarum were isolated from chickens showed typical clinical signs related with infection of pullorum disease or fowl typhoid at commercial chicken farms. DNA markers of these strains produced by GEN 70-04, GEN 70-05 and GEN 70-08 showed significant difference of band patterns between S pullorum and S gallinarum. These DNA markers could be used for comparison of DNA marker polymorphism between S pullorum and S gallinarum as well as rapid diagnosis of fowl typhoid and pullorum disease of domestic fowls.

  • PDF

Genetic Analysis study of Sasang Constitution Classification by DNA-fingerprinting methods (유전자지문법을 이용한 사상체질의 유전적 분석 연구)

  • Cho, DongWuk;Lee, ChangSoo;Ko, ByungHee;Cho, HwangSung
    • Journal of Sasang Constitutional Medicine
    • /
    • v.8 no.2
    • /
    • pp.151-163
    • /
    • 1996
  • VNTR and STR DNA typing are typical genetic analysis methods which are widely used in DNA-fingerprinting for forensic science and other genetic research purposes. In this study, genomic DNA of different constitutions(Taeun, Soyang and Soum) were analyzed by VNTR and STR DNA typing to provide scientific and objective references for Sasang Medicine. It was found out in this study that VNTR-MCT118 and YNZ22 loci showed too many different variation of allele distribution and numbers for each constitution. Therefore, it is thought that VNTR typing can not used for genetic classification study for Sasang Constitution which classifies human body into 4 groups. However, vWA locus, one of the STR loci investigated in this study, showed slight difference in allele distribution for each different constitution.

  • PDF

Analysis of Genetic Characteristics of Korean Native Chicken Using DNA Marker (DNA Marker를 이용한 한국 재래닭의 유전특성 분석)

  • 이학교;이성진;황규춘;정일정;박용호;손시환;신영수;오봉국;한재용
    • Korean Journal of Poultry Science
    • /
    • v.23 no.4
    • /
    • pp.177-183
    • /
    • 1996
  • This study was conducted to analyze genetic characteristics of Korean Native Chicken three lines classified on the basis of the feather color and appearance (Red, Yellow, and Black) using DNA fingerprinting method. To estimate the genetic relatedness among breeds and similarities within breeds, we collected blood samples from Korean Native Chicken (KNC), Rhode Island Red (RIR), White Leghorn (WL), and Cornish(CN) and obtained genomic DNA from the blood of 10 individuals randomly selected within the breeds and lines. The genomic DNA samples were digested with restriction enzymes (Hinf J, Hae Ill) and hybridized with various probes (Jeffreys' probes 33.15, 33.6 and M13) after Southern transfer. Genetic similarities within breeds were characterized by band sharing (BS) value, estimated by the DFP band pattern between the pair of lanes. BS values within WL, RIR, and KNC were 0.82, 0.70 and 0.56, respectively. Relative genetic diversity (BS value) of KNC was higher than those two breeds (WL, RIR). Estimation of genetic similarity between KNC lines and control breed (RIR) was 0.32, whereas similarity within KNC lines (6 groups) was 0.50. In this analysis, KNC was showed to have a highly genetic diver-sity at the DNA level, and to be closer in genetic distance to RIR (0.67) than any other breeds.

  • PDF

DNA Fingerprinting Analysis of Natural Monument Gingko Trees Using Microsatellite Markers (Microsatellite 마커를 이용한 은행나무 천연기념물의 DNA 지문 분석)

  • Lee, Jei-Wan;Lee, Min-Woo;Ahn, Ji-Young;Hong, Kyung-Nak;Baek, Seung-Hoon;Kim, Sang-Chul
    • Journal of Korean Society of Forest Science
    • /
    • v.106 no.4
    • /
    • pp.408-416
    • /
    • 2017
  • This study describes DNA fingerprinting analysis of twenty-three natural monument individuals of Ginkgo biloba using eight microsatellite markers. The average number of observed alleles was 6.875, and the expected heterozygosity and the observed heterozygosity were 0.711 and 0.710, respectively. This results were similar to those of the previous studies on Ginkgo trees analyzed by same markers in China and Japan. PIC value and PD were calculated at 0.677 and 0.9999 respectively, indicating a high individual identification efficiency. In fact, all of the natural monument ginkgo trees and additionally analyzed thirteen general ginkgo tress were identified by genotype comparison. PI and PD calculated in three markers (Ging06, Gb60, Gb61) with the highest PIC values calculated in natural monument ginkgo trees were $8.045{\times}10^{-5}$ and 99.99%, respectively. Thus, these three markers could be preferentially used in DNA fingerprinting for identifying ginkgo tree individuals. The results in this study will be useful for management of natural monument ginkgo trees, proliferation of their progeny and genetic identification of individuals selected in breeding process.

Studies on DNA Fingerprint for the Korean Native Chicken (한국 재래계의 유전자 지문에 관한 연구)

  • 여정수;정태완;한재용;최창본;김재우;정선부
    • Korean Journal of Poultry Science
    • /
    • v.20 no.4
    • /
    • pp.209-216
    • /
    • 1993
  • This study was conducted to classify Korean native chicken(KNC) and imported chicken by phenotypic performances and DNA fingerprinting. Two lines, KNC and White Leghorn(WL) , of chicken were maintained in the laboratory of Yeungnam University. Economic traits (body weight, sexual maturity, hen-day egg production, egg weight) and phenotypic characteristics (body-type, head, feather, shank) were checked. The DNA fingerprinting was analyzed for both breeds. The growth rate of the KNC was similar to WLS and sexual maturity of the KNC came later than WL. Hen-day egg production of the KNC was also slightly lower than the WL. The egg weight was about 10g lighter than WL. There was no difference in body weight of female KNC compared to the WL after 28 weeks. The study confirms difference between KNC and WL in DNA fingerprinting as well as its outlook. Thus, we suggest that these should be tested in nationwide districts about chickens known as the KNC using DNA fingerprinting. Then, the confirmed KNC populations should be maintained and used for the genetic improvement. Finally, only confirmed KNC should be in market which induce consumer to seek the KNC by its favorite.

  • PDF

Genotypic and Phenotypic Diversity of PGPR Fluorescent Pseudomonads Isolated from the Rhizosphere of Sugarcane (Saccharum officinarum L.)

  • Rameshkumar, Neelamegam;Ayyadurai, Niraikulam;Kayalvizhi, Nagarajan;Gunasekaran, Paramsamy
    • Journal of Microbiology and Biotechnology
    • /
    • v.22 no.1
    • /
    • pp.13-24
    • /
    • 2012
  • The genetic diversity of plant growth-promoting rhizobacterial (PGPR) fluorescent pseudomonads associated with the sugarcane (Saccharum officinarum L.) rhizosphere was analyzed. Selected isolates were screened for plant growthpromoting properties including production of indole acetic acid, phosphate solubilization, denitrification ability, and production of antifungal metabolites. Furthermore, 16S rDNA sequence analysis was performed to identify and differentiate these isolates. Based on 16S rDNA sequence similarity, the isolates were designated as Pseudomonas plecoglossicida, P. fluorescens, P. libaniensis, and P. aeruginosa. Differentiation of isolates belonging to the same group was achieved through different genomic DNA fingerprinting techniques, including randomly amplified polymorphic DNA (RAPD), amplified ribosomal DNA restriction analysis (ARDRA), repetitive extragenic palindromic (REP), enterobacterial repetitive intergenic consensus (ERIC), and bacterial repetitive BOX elements (BOX) analyses. The genetic diversity observed among the isolates and rep-PCR-generated fingerprinting patterns revealed that PGPR fluorescent pseudomonads are associated with the rhizosphere of sugarcane and that P. plecoglossicida is a dominant species. The knowledge obtained herein regarding the genetic and functional diversity of fluorescent pseudomonads associated with the sugarcane rhizosphere is useful for understanding their ecological role and potential utilization in sustainable agriculture.