• Title/Summary/Keyword: RFLP patterns

Search Result 143, Processing Time 0.02 seconds

Toll-like Receptor 4 Polymorphism and Periodontitis in Korean Population

  • Park, Ok-Jin;Shin, Seung-Yun;Chung, Chong-Pyoung;Ku, Young;Choi, Young-Nim;Kim, Kack-Kyun
    • International Journal of Oral Biology
    • /
    • v.31 no.1
    • /
    • pp.1-6
    • /
    • 2006
  • The primary cause of periodontitis is plaque-associated anaerobic gram-negative bacteria. As shown in the patients with defects in the number or function of neutrophils, innate immunity plays an important role in resistance to bacterial infection and periodontitis. Toll-like receptor 4(TLR4) is one of the key receptors that recognize the molecular patterns of microbes and initiate innate immune response. To understand the role of TLR4 in the pathogenesis of periodontitis, we investigated whether Asp299Gly of TLR4 mutation is associated with periodontitis in Korean population. Subjects for this study included 90 healthy subjects and 98 periodontitis patients. The Asp299Gly mutation was screened by PCR-Restriction Fragment Length Polymorphism(RFLP) of genomic DNA from blood cells using a primer that creates a NcoI restriction site only in the mutant allele. The Asp299Gly mutation was not found in all subjects tested. Our results suggest that the Asp299Gly mutation of TLR4 is very rare in a Korean population. Further mutation screening may be required to determine the role of TLR4 in the pathogenesis of periodontitis.

Genotypic Identification of Cystoisospora in Immunocompromised Patients Using Tm-Variation Analysis

  • Basyoni, Maha M.A.;Elghobary, Hany Ahmed Fouad
    • Parasites, Hosts and Diseases
    • /
    • v.55 no.6
    • /
    • pp.601-606
    • /
    • 2017
  • Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR), and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures ($T_ms$) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified $T_ms$ at $85.8^{\circ}C$ and $88.6^{\circ}C$, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using $qPCR-T_ms$ analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups.

Pulsed-Field Gel Electrophoresis and Mutation Typing of gyrA Gene of Quinolone-Resistant Salmonella enterica Serovar Paratyphi A Isolated from Outbreak and Sporadic Cases,1998-2002, Korea

  • KIM SHUKHO;OK YOUNG LIM;SEONG HAN KIM;JUN YOUNG KIM;YEON HO KANG;BOK KWON LEE
    • Journal of Microbiology and Biotechnology
    • /
    • v.13 no.1
    • /
    • pp.155-158
    • /
    • 2003
  • In early 2002, over 200 people in the city of Pusan. Korea suffered from paratyphoid fever resulting from Salmonella Paratyphi A Infection. Antimicrobial susceptibility tests and Xbal pulsed-field gel electrophoresis (PFCE) were conducted to 54 Salmonella Paratyphi A isolated from humans during the period of 1998 to 2002. Most of the isolates ($83\%$) were only nalidixic acid-resistant and $78\%$ were X 1 PFGE patterns. Also, we measured the MIC of ciprofloxacin and screened gyrA mutation(5) using allele- specific PCR and restriction fragment length polymorphism (AS-PCR-RFLP). The representative 5 isolates in 2002 and 1 isolate in 2000 were $1{\mu}g/ml$ of MIC and had mutation at the 83rd codon in gyrA. These data suggest that the outbreak in the early 2002 might have been due to dissemination of the strain present In 2000. Also, decreased susceptibility to ciprofloxacin was partly due to the mutation at the 83rd codon in gyrA.

Urinary Bladder Cancer Risk Factors: A Lebanese Case-Control Study

  • Kobeissi, Loulou Hassan;Yassine, Ibrahim Adnan;Jabbour, Michel Elias;Moussa, Mohamad Ahmad;Dhaini, Hassan Rida
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.5
    • /
    • pp.3205-3211
    • /
    • 2013
  • Background: Bladder cancer is the second most incident malignancy among Lebanese men. The purpose of this study was to investigate potential risk factors associated with this observed high incidence. Methods: A case-control study (54 cases and 105 hospital-based controls) was conducted in two major hospitals in Beirut. Cases were randomly selected from patients diagnosed in the period of 2002-2008. Controls were conveniently selected from the same settings. Data were collected using interview questionnaire and blood analysis. Exposure data were collected using a structured face-to-face interview questionnaire. Blood samples were collected to determine N-acetyltransferase1 (NAT1) genotype by PCR-RFLP. Analyses revolved around univariate, bivariate and multivariate logistic regression, along with checks for effect modification. Results: The odds of having bladder cancer among smokers was 1.02 times significantly higher in cases vs. controls. The odds of exposure to occupational diesel or fuel combustion fumes were 4.1 times significantly higher in cases vs controls. The odds of prostate-related morbidity were 5.6 times significantly higher in cases vs controls. Cases and controls showed different clustering patterns of NAT1 alleles. No significant differences between cases and controls were found for consumption of alcohol, coffee, tea, or artificial sweeteners. Conclusions: This is the first case-control study investigating bladder cancer risk factors in the Lebanese context. Results confirmed established risk factors in the literature, particularly smoking and occupational exposure to diesel. The herein observed associations should be used to develop appropriate prevention policies and intervention strategies, in order to control this alarming disease in Lebanon.

Genetic Analysis of Mitochondrial DNA from Korean Oysters, Crassostrea gigas (한국산 참굴(Crassostrea gigas) 미토콘드리아 DNA의 유전적 분석)

  • KIM Sang Hae;PARK Mi Seon;KIM Young Hun;PARK Doo Won
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.30 no.5
    • /
    • pp.804-808
    • /
    • 1997
  • The genetic differentiation and characteristics of two oyster populations (Crassostrea gigas) in Korea were assessed based on the restriction fragment length polymorphisms (RFLP) analysis and the restriction patterns of subcloned mtDNA. The restriction fragments of twenty individuals in West Sea revealed an identical pattern, determined by 8 restriction enzymes. On the other hand, two haplotypes having variation at the HindIII site were shown in the specimens from South Sea; minor haplotypes (4 of 20) were similar to the results obtained from individuals in West Sea while major haplotypes were different from those in West Sea. It was suggested that oysters (C. gigas) of West Sea might have been introduced to South Sea. Each mitochondrial DNA from two oyster populations in Korea and from one in Japan was divided to three parts and subcloned into pUC19 to use in genetic studies effectively. Restriction map was constructed based on the cleavage pattern by multiple restriction enzymes.

  • PDF

Association Between MSTN Gene Polymorphism and Growth Traits in Landrace Pigs (돼지 Landrace 품종에서 Myostatin 유전자의 유전적 다형성과 성장형질과의 연관성)

  • Cho, I.C.;Choi, Y.L.;Ko, M.S.;Kim, H.S.;Lee, J.G.;Jeon, J.T.;Han, Sang-Hyun
    • Journal of Animal Science and Technology
    • /
    • v.47 no.2
    • /
    • pp.159-166
    • /
    • 2005
  • Porcine myostatin(MS1N) gene plays a key role in the differentiation of myoblast and muscle development. Genetic polymorphism was screened by single stranded conformation polymorphism(SSCP) analysis and subsequent DNA sequencing detected a nucleotide substitution(C2150T) in exon 3 of MSIN gene. Phenotypic association of the polymorphism was tested in a Landrace population and positive effects of the allele T for lean growth traits were found in the population. Even though it is not significant, the pigs have IT and TC genotypes were heavier for the body weight at birth and at twenty weeks of age than those containing genotype. Cc. However, the allele T was significantly associated with higher eye muscle area(P < 0.05). As a result of this study, we suggested that the allele T in exon 3 of MSTN gene comes a significant effect for increasing the eye muscle area without decreasing backfat thickness. This polymorphism did not change the amino acid but Taq I -RFLP matched to SSCP band patterns in exon 3 of MSTN gene, which will be an useful molecular marker for breeding of Landrace pigs.

First Note of Hypoxylon truncatum sensu Miller in Korea (국내에서 발견된 Hypoxylon truncatum sensu Miller의 형태 및 유전적 특징)

  • Yoon, Cheol-Sik;Chang, Hyun-You;Sung, Gi-Ho
    • The Korean Journal of Mycology
    • /
    • v.25 no.3 s.82
    • /
    • pp.161-166
    • /
    • 1997
  • Occurrence of Hypoxylon truncatum, known as a symbiont of white jelly mushroom, was first noted and described in Korea. Two stromatal forms reported in North American collections of H. truncatum sensu Miller were also observed in Korean collections. Based on evidences from other studies, polymorphic patterns seen in six enzyme digestions of nuclear Internal Transcribed Spacer (ITS) regions of Korean isolates indicated that the two stromatal forms were genetically distinct. Because there was a clear association of stromatal morphology with genetic differences, the different stromatal forms might be different species. In addition, clear species concept on the species H. truncatum would provide aids in selecting proper strain for cultivation of white jelly mushrooms.

  • PDF

Relationship between Stress Gene Polymorphisms and Litter Size by AI in Pigs

  • Jin, H.J.;Kim, I.C.;Wee, M.S.;Yeon, S.H.;Kim, C.D.;Lee, S.S.;Cho, C.Y.;Cho, S.R.;Son, D.S.;Park, C.K.;Li, Z.D.
    • Journal of Embryo Transfer
    • /
    • v.22 no.4
    • /
    • pp.257-263
    • /
    • 2007
  • This study was performed to investigate the relationship between PSS-HSP70 gene polymorphism and artificial insemination (AI) reproductivity in the pigs. The RFLP polymorphism of PSS and the SSCP polymorphisms of HSP70 K1, K3 and K4 PCR product were detected different patterns. In the experiment for AI of fresh semen, spring and fall season showed higher litter size born of 10.89 head than 10.47 head of summer season. Landrace was showed higher litter size of 9.96 head than that of Duroc and Yorkshire (p<0.05). Stress relating PSS and HSP70 polymorphism of PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showd a highest litter size born of 10.97 head and litter size born alive of 10.69 head than that of the other polymorphisms(p<0.05). In the experiment for AI of frozen semen, effects of season and pig breeds were not showed for litter size born. The stress relating polymorphism of PSS-Carrier, HSP70 K1-BB, K3-BB, K4-AB showed highest litter size born of 11.29 head and litter size born alive of 10.82 head and PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showed the lowest litter size born of 8.48 head and litter size born alive of 7.33 head than that of the other polymorphisms(p<0.05). These results suggest that AI litter size born for the stress of forzen thawed semen may be affected by PSS and HSP70 polymorphism in pigs.

Characterization of a Novel cry1-Type Gene from Bacillus thuringiensis subsp. alesti Strain LY-99

  • Qi, Xu Feng;Li, Ming Shun;Choi, Jae-Young;Roh, Jong-Yul;Song, Ji Zhen;Wang, Yong;Jin, Byung-Rae;Je, Yeon-Ho;Li, Jian Hong
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.18 no.1
    • /
    • pp.18-27
    • /
    • 2009
  • B. thuringiensis strain LY-99 belonging to subsp. alesti (H3a3c), was isolated from Chinese tobacco warehouse and showed significantly high toxicity to Plutella xylostella. For the identification of the cry1-type genes from B. thuringiensis LY-99, an extended multiplex PCRrestriction fragment length polymorphism (PCRRFLP) method was established by using two pairs of universal primers based on the conserved regions of the cry1-type genes to amplify around 2.4 kb cry1-type gene fragments. Then the DNA fragment was cloned into pGEM-T Easy vector and digested with EcoRI and EcoRV enzymes. Through this method, a known cry1-type gene was successfully identified from the reference strain, B. thuringiensis subsp. alesti. In addition, the RFLP patterns revealed that B. thuringiensis LY-99 included a novel cry1A-type gene in addition to cry1Aa, cry1Ac, cry1Be and cry1Ea genes. The novel cry1A-type gene was designated cry1Ah2 (Genbank accession No DQ269474). An inverse PCR method was used to amplify the flank regions of cry1Ah2 gene. Finally, 3143 bp HindIII fragment from B. thuringiensis LY-99 plasmid DNA including 5' region and partial ORF was amplified, and sequence analysis revealed that cry1Ah2 gene from LY-99 showed 89.31% of maximum sequence similarity with cry1Ac1 crystal protein gene. In addition, the deduced amino acid sequence of Cry1Ah2 protein shared 87.80% of maximum identity with that of Cry1Ac2. This protein therefore belongs to a new class of B. thuringiensis crystal proteins.

Studies on the Possible Relationship of Porcine Serum Insulin-like Growth Factor-I with Litter Size (돼지의 혈청 Insulin-like Growth Factor-I과 산자수간의 연관성 연구)

  • Yang, S.H.;Seo, D.S.;Park, H.B.;Kim, K.D.;Kang, C.W.;Choi, K.S.;Park, S.S.;Hong, K.C.;Ko, Y.
    • Korean Journal of Animal Reproduction
    • /
    • v.23 no.3
    • /
    • pp.213-220
    • /
    • 1999
  • The litter size has been the primary interest of economic traits in pig reproduction. It has been recently shown that insulin-like growth factor-Ⅰ(IGF-Ⅰ) plays roles in establishing pregnancy and in supporting fetal growth and development. But, the effect of serum IGF-Ⅰ on litter size has not been studied. Therefore, this study was conducted to relate serum IGF-Ⅰ concentration with porcine litter size and to investigate the possible connection with estrogen receptor(ER) as a candidate gene for the litter size. Sera during day 45 to 105 of pregnancy were collected from two groups showing high and low litter size and serum IGF-Ⅰ concentration was measured by radioimmunoassay (RIA). IGF-Ⅰ levels in both groups decreased gradually as pregnancy stage proceeded but were not significantly different. Secondly, DNA was extracted from blood and PCR-RFLP was utilized to analyze ER genotypes of pigs in each group, which produced three polymorphic patterns. Based on the ER genotypes analyzed, low litter size group showed higher IGF-Ⅰ concentration than high litter size group. Taken together, the results indicate that the serum IGF system was correlated with steroid system but not with the litter size in pigs. Thus, this study implies that porcine litter size could be determined locally at the ovary level.

  • PDF