• Title/Summary/Keyword: Genetic screening

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Strain Improvement and Genetic Characterization of Tautomycetin Biosynthesis in Streptomyces spp.

  • Choi, Si-Sun;Kim, Myung-Gun;Kim, Eung-Soo
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.420-422
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    • 2005
  • TMC (Tautomycetin) is a liner polyketide immunosuppressive antifungal compound produced by Streptomyces spp. Inhibition of T cell proliferation with TMC was observed highly efficient at 100-fold lower than those needed to achieve maximal inhibition with cyclosporin A. To elucidate the biosynthetic pathway of TMC, a genomic DNA library was constructed using a E. coil-Streptomyces shuttle cosmid vector, pOJ446. The DNA libraries were screened by colony blot hybridization using several polyketide ${\beta}-ketosynthase$ (KS) probes amplified from TMC-producing Streptomyces genomic DNA using polymerase chain reaction (PCR), of which the degenerate primers were designed based on the highly conserved sequences present in KS domains of various type I polyketide synthase genes in Streptomyces species. This library construction and screening approach led to the isolation of several positive cosmid clones representing type I polyketide biosynthetic gene clusters. In addition, a Streptomyces regulatory gene called afsR2 (a global regulatory gene stimulating antibiotic production in both S. coelicolor and S. lividans) was successfully integrated into the TMC-producing Streptomyces chromosome via E. coil-Streptomyces heterologous conjugation mehtod. The more detailed results of production improvement and genetic characterization of TMC-producing Streptomyces spp. will be discussed.

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Antibiotics produced by anaerobic fermentation of Streptococcus sp. An-21-1 isolated from domestic soil I. Screening and identification of anaerobic bacteria (국내토양에서 분리한 혐기성 세균 Streptococcus sp. An-21-1 이 생성하는 항생물질 I. 혐기성 세균의 선별과 동정)

  • Park, Seung-chun;Yun, Hyo-in;Oh, Tae-kwang
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.53-60
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    • 1993
  • Anaerobic bacteria are suggested to be potential source for new antibiotics. In order to search for antibiotics from domestic origin, we collected 800 soil samples across Korean locations and could isolate as many as 989 anaerobic strains. Among them 10, strains were found to have good producing capacity of antibiotics. An anaerobe was finally selected due to secreting antibiotics having high antimicrobial activity towards multiple resistant microorganism(E coli JM 83) transformed by genetic engineering technique. Its morphological, physiological and biochemical charateristics were investigated, together with antimicrobial spectrum therefrom. On antimicrobial spectrum study, substance secreted from this strain, had no activities to fungus and yeast. The selected strain showed G(+) and coccal shape, on Gram, staining and electron scanning microscopy, respectively. Biochemically this strain utilized glucose, fructose lactose, sucrose, but did not arabinose, cellulose, rhamnose, sorbitol, trehalose, mannitol. Catalase test showed negative property. Optimal growth temperature was $37^{\circ}C$. The results obtained above suggest this strain Streptococcus faecium subspp. and we named it Streptococcus sp. An-21-1.

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Genetic Variation in a DNA Double Strand Break Repair Gene in Saudi Population: A Comparative Study with Worldwide Ethnic Groups

  • Areeshi, Mohammed Yahya
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7091-7094
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    • 2013
  • DNA repair capacity is crucial in maintaining cellular functions and homeostasis. However, it can be altered based on DNA sequence variations in DNA repair genes and this may lead to the development of many diseases including malignancies. Identification of genetic polymorphisms responsible for reduced DNA repair capacity is necessary for better prevention. Homologous recombination (HR), a major double strand break repair pathway, plays a critical role in maintaining the genome stability. The present study was performed to determine the frequency of the HR gene XRCC3 Exon 7 (C18067T, rs861539) polymorphisms in Saudi Arabian population in comparison with epidemiological studies by "MEDLINE" search to equate with global populations. The variant allelic (T) frequency of XRCC3 (C>T) was found to be 39%. Our results suggest that frequency of XRCC3 (C>T) DNA repair gene exhibits distinctive patterns compared with the Saudi Arabian population and this might be attributed to ethnic variation. The present findings may help in high-risk screening of humans exposed to environmental carcinogens and cancer predisposition in different ethnic groups.

Comparison of Multilocus Sequence Typing (MLST) and Repetitive Sequence-Based PCR (rep-PCR) Fingerprinting for Differentiation of Campylobacter jejuni Isolated from Broiler in Chiang Mai, Thailand

  • Patchanee, Prapas;Chokboonmongkol, Chomporn;Zessin, Karl-Hans;Alter, Thomas;Pornaem, Sarinya;Chokesajjawatee, Nipa
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1467-1470
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    • 2012
  • We compared rapid fingerprinting using repetitive sequencebased PCR (rep-PCR) for subtyping Campylobacter jejuni isolates to the widely used multilocus sequence typing (MLST). Representative C. jejuni isolates (n = 16) from broilers were analyzed using MLST and rep-PCR. Both techniques demonstrated an equal discriminatory power of 0.8917, and 9 subgroups were identified. Clonal identification of all 16 isolates was identical for both techniques. The rep-PCR as described in this study may be used as a rapid and cost-effective alternative for subtyping of C. jejuni isolates, or as an effective screening tool in large epidemiological studies.

DNA testing for fragile X syndrome in school for severely emotionally handicapped children in Korea

  • Hong, Sung-Do David;Lee, So-Young;Oh, Myung-Ryurl;Jin, Dong-Kyu
    • Journal of Genetic Medicine
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    • v.2 no.2
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    • pp.83-86
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    • 1998
  • Though Fragile X syndrome is one of the most common inherited causes of mental retardation, it is not much detected yet in Korean population. One of the reason may be that the syndrome is not well known to the special education teachers as well as to the clinicians in this country. Thus, molecular test was undertaken to screen out fragile X syndrome in 122 children of two Korean schools for emotionally severely handicapped children. The subjects were all boys, previously known as having pervasive developmental disorder with or without mental retardation. Southern blot analysis of peripheral blood showed the abnormally enlarged (CGG)n repeat sequence associated with fragile X syndrome in two children. This finding suggests that the DNA testing for fragile X syndrome is warranted for Korean high risk population and that more concern about this syndrome is needed for the professionals who work for mentally handicapped children. The issues involved in genetic counseling for fragile X syndrome are discussed.

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Screening for Del 185 AG and 4627C>A BRCA1 Mutations in Breast Cancer Patients from Lahore, Pakistan

  • Aziz, Faiza;Fatima, Warda;Mahmood, Saqib;Khokher, Samina
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1725-1727
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    • 2016
  • Breast cancer contributes to approximately 23% of the cancer cases identified and 14% of cancer related deaths worldwide. Including a strong association between genetic and environmental factors, breast cancer is a complex and multi factorial disorder. Two high penetration breast cancer susceptibility genes (BRCA1 and BRCA2) have been identified, and germ line mutations in these are thought to account for between 5% and 10% of all breast cancer cases. The human BRCA1 gene, located on 17q, is involved in the regulation of cell proliferation by aiding in DNA repair, transcriptional responses to DNA damage and cell cycle check points. Mutations in this gene enhance cell proliferation and facilitate formation of tumors. Two mutations, the 185 deletion of AG and the 4627 substitution from C to A, are founder mutations in the BRCA1 gene for breast cancer in Asian populations. Allele specific PCR was performed to detect these selected mutations in 120 samples. No mutation of 4627 C to A was detected in the samples and only one of the patients had the 185 del AG mutation in the heterozygous condition. Our collected samples had lower consanguinity and family history indicating the greater involvement of environmental as compared to genetic factors.

Biomonitoring the Genotoxicity of Environmental Pollutants Using the Tradescantia Bioassay (환경 중 유전독성물질 검색을 위한 자주달개비 생물검정 기법의 적용연구)

  • 신해식
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2004.05a
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    • pp.47-60
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    • 2004
  • Higher plants can be valuable genetic assay systems for monitoring environmental pollutants and evaluating their biological toxicity. Two assays are considered ideal for in situ monitoring and testing of soil, airborne and aqueous mutagenic agents; the Tradescantia stamen hair assay for somatic cell mutations and the Tradescantia micronucleus assay for chromosome aberrations. Both assays can be used for in vivo and in vitro testing of mutagens. Since higher plant systems are now recognized as excellent indicators and have unique advantages over in situ monitoring and screening, higher plant systems could be accepted by regulatory authorities as an alternative first-tier assay system for the detection of possible genetic damages resulting from the pollutants or chemicals used and produced by industrial sectors. It has been concluded that potential mutagen and carcinogen such as the heavy metals among indoor air particulates, volatile compounds in the working places, soil, and water pollutants contribute to the overall health risk. This contribution can be considerable under certain circumstances. It is therefore important to identify the level of genotoxic activity in the environment and to relate it to the biomarkers of a health risk in humans. The results from the higher plant bioassays could make a significant contribution to assessing the risks of pollutants and protecting the public from agents that can cause mutation and/or cancer. The plant bioassays, which are relatively inexpensive and easy to handle, are recommended for the scientists who are interested in monitoring pollutants and evaluating their environmental toxicity to living organisms.

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Assessment of Environmental Pollution with Tradescantia Bioassays (자주달개비 생물검정 기법을 이용한 환경오염 평가)

  • Kim Jin Gyu;Sin Hae Sik
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2004.05a
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    • pp.1-15
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    • 2004
  • Higher plants can be valuable genetic assay systems for monitoring environmental pollutants and evaluating their biological toxicity. Two assays are considered ideal for in situ monitoring and testing of soil, airborne and aqueous mutagenic agents; the Tradescantia stamen hair assay for somatic cell mutations and the Tradescantia micronucleus assay for chromosome aberrations. Both assays can be used for in vivo and in vitro testing of mutagens. Since higher plant systems are now recognized as excellent indicators and have unique advantages over in situ monitoring and screening, higher plant systems could be accepted by regulatory authorities as an alternative first-tier assay system for the detection of possible genetic damages resulting from the pollutants or chemicals used and produced by industrial sectors. It has been concluded that potential mutagen and carcinogen such as the heavy metals among indoor air particulates, volatile compounds in the working places, soil, and water pollutants contribute to the overall health risk. This contribution can be considerable under certain circumstances. It is therefore important to identify the level of genotoxic activity in the environment and to relate it to the biomarkers of a health risk in humans. The results from the higher plant bioassays could make a significant contribution to assessing the risks of pollutants and protecting the public firom agents that can cause mutation anuor cancer. The plant bioassays, which are relatively inexpensive and easy to handle, are recommended for the scientists who are interested in monitoring pollutants and evaluating their environmental toxicity to living organisms.

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Biological Screening of a Novel Nickel (II) Tyrosine Complex

  • Islam, Md. Rafiqul;Islam, S.M. Rafiqul;Noman, Abu Shadat Mohammod;Khanam, Jahan Ara;Ali, Shaikh Mohammad Mohsin;Alam, Shahidul;Lee, Min-Woong
    • Mycobiology
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    • v.35 no.1
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    • pp.25-29
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    • 2007
  • A newly synthesized Nickel (II) tyrosine complex was screened as potential antimicrobial agent against a number of medically important bacteria (Bacillus subtilis, Streptococcus ${\beta}$-haemolytica, Escherichia coli, Shigella dysenterae) and fungi (Aspergillus fumigatus, Candida albicans, Aspergillus niger, Aspergillus flavus, Penicillium sp.) strains. were used for antifungal activity. The antimicrobial activity was evaluated using the Agar Disc method. Moreover, the minimum inhibitory concentration of the complexes was determined against the same pathogenic bacteria and the values were found between $4{\sim}64\;{\mu}g\;ml^{-1}$. Brine shrimp bioassay was carried out for cytotoxicity measurements of the complexes. The $LC_{50}$ values were calculated after probit transformation of the resulting mortality data and found to be 6 ${\mu}g\;ml^{-1}$.

A Case of Galactosemia with Novel Mutation in the GALT Gene (새로운 GALT 유전자의 돌연변이에 의한 갈락토스혈증)

  • Kim, Shin Ah;Shin, Young Lim;Hong, Yong Hee
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.13 no.2
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    • pp.126-130
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    • 2013
  • Galactosemia is a metabolic disorder inherited by the recessive autosome, and appears by the deficiency of one enzyme out of GALT (Galactose-1-Phosphate Uridyltransferase), GALK (galactokinase), and GALE (epimerase) enzymes, among which the GALT deficiency disease is denominated as classical galactosemia and known to have symptoms such as severe nausea, jaundice, hepatomegaly, sucking difficulty and so on. We report the case of a 16-day-old female baby with the new p.A101D mutation together with p.N413d in the GALT gene analysis found in the neonatal screening test and diagnosed to have galactosemia by the GALT deficiency through the enzyme analysis. For the prognosis prediction, the treatment, the genetic counseling and the prenatal diagnosis of the patients, more detailed genetic diagnosis is required by performing GALT gene analysis, and it is deemed to be necessary to analyze the correlation between the phenotype and the genotype of the domestic galactosemia patients.

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