• Title/Summary/Keyword: specific probe

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유전공학기법으로 변형시킨 내성유전자네 대한 수질환경에서의 전이동태

  • 이성기;김치경
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.322-331
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    • 1992
  • In order to understand the transfer and behavior of R gene in water environments. the Kmr gene in the genetically modified microorganisms(GMMs) w,is studied by conjugation. The plasmid variously rearranged in the conjugants were comparatively analyzied by agarosc gel electrophoresis and the specific Km' genes in the gel were tletected with DNA probe. The Kmr genes of the GMM strains(DKC600 and DKC601) were transferred at higher rate than those of natural isola~e(DKI)b, ut the ratc was a little diflurent depending upon the recipient strains. Rearrangement of the plasmids appeared morc drastic in GMM strains than in IIKI as donor. The transfer frequencies of the Km' genes in LR broth were remarkably higher than in the water of AW and FW without regards to the strains. In LA breth. the frequencies of Kmr genes were higher at 25'C-30$^{\circ}$C than at 10$^{\circ}$C and at pH - 7 than pH 9, but temperature and pH of the FW did n,,t affect to the frequency. And the conjugants from GMM strains in FW did not showed any plasmids. except tor 43 kb plasmiil. As results of Southern analysis of the plasmid, variously rearranged in eonjugant cells obtained in LB broth, the Kmr genes were detected at the same position of Km' plasrnids of the donor cell(DK1 and GMM strains). But Km' plasmid disappeared in the conjugants obtained in F'W and their chronlosomes showed strong signal of hybridization. The Kmr plasmid of DKl in the conjugants obtained in FW water was transferred and maintained its size, but the Kmr plasinids of the GMM strains were all integrated into chromosome. Therefore, the Kmr plasmids of DKI anit GMM strains in LH were intactly transferred and other plasmitls were variously rearranged. but Km' gene of DKC600 in FW water was integrated into the chromosorn: without regards to the temperature and pH of the water.

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Detection of Point Mutations in the rpoB Gene Related to Drug Susceptibility in Mycobacterium Tuberculosis using an Oligonucleotide Chip (올리고뉴클레오티드 칩(Oligonucleotide Chip)을 이용한 항결핵제 감수성과 관련된 Mycobacterium tuberculosis rpoB 유전자의 점돌연변이 판별 방법)

  • Kim, Hyun-Jung;Kim, Seong-Keun;Shim, Tae-Sun;Park, Yong-Doo;Park, Mi-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.29-41
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    • 2001
  • Background : The appearance of multiple-drug-resistant Mycobacterium tuberculosis strains has been seriously compromising successful control of tuberculosis. Rifampin-resistance, caused by mutations in the rpoB gene, can be indicative of multiple-drug-resistance, and its detection is of great importance. The present study aimed to develop an oligonucleotide chip for accurate and convenient screening of drug-resistance. Methods : In order to detect point mutations in the rpoB gene, an oligonucleotide chip was prepared by immobilizing specific probe DNA to a microscopic slide glass by a chemical reaction. The probe DNA that was selected from the 81 bp core region of the rpoB gene was designed to have mutation sites at the center. A total of 17 mutant probes related to rifampin-resistance including 8 rifabutin-sensitive mutant probes were used in this study. For accurate determination, wild type probes were prepared for each mutation position with an equal length, which enabled a direct comparison of the hybridization intensities between the mutant and wild type. Results : Mycobacterial genomic DNA from clinical samples was tested with the oligonucleotide chip and the results were compared with those of the drug-susceptibility test in addition to sequencing and INNO-LiPA Rif. TB kit test in some cases. Out of 15 samples, the oligonucleotide chip results of 13 samples showed good agreement with the rifabutin-sensitivity results. The two samples with conflicting result also showed a discrepancy between the other tests, suggesting such possibilities as existence of mixed strains and difference in drug-sensitivity. Further verification of these samples in addition to more case studies are required before the final evaluation of the oligonucleotide chip can be made. Conlcusion : An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drugsusceptibility. The results to date show the potential for using the oligonucleotide chip for accurate and convenient screening of drug-resistance to provide useful information in antituberculosis drug therapy.

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Synthesis and Preliminary Evaluation of $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$ Guanine $([^{18}F]FHBG)$ in HSV1-tk Gene Transduced Hepatoma Cell (9-(4-$[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine $([^{18}F]FHBG)$의 합성과 헤르페스 단순 바이러스 티미딘 키나아제 이입 간암 세포주에서의 기초 연구)

  • Moon, Byung-Seok;Lee, Tae-Sup;Lee, Myoung-Keun;Lee, Kyo-Chul;An, Gwang-Il;Chun, Kwon-Soo;Awh, Ok-Doo;Chi, Dae-Yoon;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.4
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    • pp.218-227
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    • 2006
  • Purpose: The HSV1-tk reporter gene system is the most widely used system because of its advantage that direct monitoring is possible without the introduction of a separate reporter gene in case of HSV1-tk suicide gene therapy. In this study, we investigate the usefulness of the reporter probe (substrate), $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine ($[^{18}F]FHBG$) for non-invasive reporter gene imaging using PET in HSV1-tk expressing hepatoma model. Materials and Methods: Radiolabeled FHBG was prepared in 8 steps from a commercially available triester. The labeling reaction was carried out by NCA nucleophilic substitution with $K[^{18}F]/K2.2.2.$ in acetonitrile using N2-monomethoxytrityl-9-14-(tosyl)-3-monomethoxytritylmethylbutyl]guanine as a precursor, followed by deprotection with 1 N HCl. Preliminary biological properties of the probe were evaluated with MCA cells and MCA-tk cells transduced with HSV1-tk reporter gene. In vitro uptake and release-out studies of $[^{18}F]FHBG$ were performed, and was analyzed correlation between $[^{18}F]FHBG$ uptake ratio according to increasing numeric count of MCA-tk cells and degree of gene expression. MicroPET scan image was obtained with MCA and MCA-tk tumor bearing Balb/c-nude mouse model. Results: $[^{18}F]FHBG$ was purified by reverse phase semi-HPLC system and collected at around 16-18 min. Radiothemical yield was about 20-25%) (corrected for decay), radiochemical purity was >95% and specific activity was around >55.5 $GBq/{\mu}\;mol$. Specific accumulation of $[^{18}F]FHBG$ was observed in HSV1-tk gene transduced MCA-tk cells but not in MCA cells, and consecutive 1 hour release-out results showed more than 86% of uptaked $[^{18}F]FHBG$ was retained inside of cells. The uptake of $[^{18}F]FHBG$ was showed a highly significant linear correlation ($R^2=0.995$) with increasing percentage of MCA-tk numeric cell count. In microPET scan images, remarkable difference of accumulation was observed for the two type of tumors. Conclusion: $[^{18}F]FHBG$ appears to be a useful as non-invasive PET imaging substrate in HSV1-tk expressing hepatoma model.

Fabrication of Multiple-Frequency Exposure System for In Vitro Experiment (세포 실험용 다중 주파수 동시 노출 장치 제작)

  • Kim, Tae-Hong;Seo, Min-Gyeong;Mun, Ji-Yeon;Pack, Jeong-Ki
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.23 no.2
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    • pp.213-219
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    • 2012
  • Recently, we are simultaneously exposed by various electromagnetic sources due to an increase of mobile communication services. However, EMF(Electric, Magnetic and Electromagnetic Field) study has been performed mainly about only single frequency. The objective of this paper is to develop an multiple-frequency exposure system for in vitro experiment. The exposure unit for in vitro experiments was designed by radial transmission line type to get broadband characteristics to generate signals of CDMA at 836.5 MHz and WCDMA at 1950 MHz frequency simultaneously. The modulated signals were delivered to the conical antenna through amplifier, digital attenuator and RF combiner. SAR values were obtained by the averaged values of 3 measured values at 9 points in petri dish using the fiber optic temperature probe. The measured return loss was under -15 dB. For 1 W input power, the mean value and standard deviation of SAR were $0.105{\pm}0.019$ for the CDMA frequency and $0.262{\pm}0.055$ for the WCDMA frequency.

Molecular Cloning and Bioinformatic Analysis of SPATA4 Gene

  • Liu, Shang-Feng;Ai, Chao;Ge, Zhong-Qi;Liu, Hai-Luo;Liu, Bo-Wen;He, Shan;Wang, Zhao
    • BMB Reports
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    • v.38 no.6
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    • pp.739-747
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    • 2005
  • Full-length cDNA sequences of four novel SPATA4 genes in chimpanzee, cow, chicken and ascidian were identified by bioinformatic analysis using mouse or human SPATA4 cDNA fragment as electronic probe. All these genes have 6 exons and have similar protein molecular weight and do not localize in sex chromosome. The mouse SPATA4 sequence is identified as significantly changed in cryptorchidism, which shares no significant homology with any known protein in swissprot databases except for the homologous genes in various vertebrates. Our searching results showed that all SPATA4 proteins have a putative conserved domain DUF1042. The percentages of putative SPATA4 protein sequence identity ranging from 30% to 99%. The high similarity was also found in 1 kb promoter regions of human, mouse and rat SPATA4 gene. The similarities of the sequences upstream of SPATA4 promoter also have a high proportion. The results of searching SymAtlas (http://symatlas.gnf.org/SymAtlas/) showed that human SPATA4 has a high expression in testis, especially in testis interstitial, leydig cell, seminiferous tubule and germ cell. Mouse SPATA4 was observed exclusively in adult mouse testis and almost no signal was detected in other tissues. The pI values of the protein are negative, ranging from 9.44 to 10.15. The subcellular location of the protein is usually in the nucleus. And the signal peptide possibilities for SPATA4 are always zero. Using the SNPs data in NCBI, we found 33 SNPs in human SPATA4 gene genomic DNA region, with the distribution of 29 SNPs in the introns. CpG island searching gives the data about CpG island, which shows that the regions of the CpG island have a high similarity with each other, though the length of the CpG island is different from each other.This research is a fundamental work in the fields of the bioinformational analysis, and also put forward a new way for the bioinformatic analysis of other genes.

Risperidone as a Janus in Mood Disorder (기분장애에서 risperidone의 양면성)

  • Yoon, Doh Joon
    • Korean Journal of Biological Psychiatry
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    • v.4 no.2
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    • pp.198-210
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    • 1997
  • To examine the double-faced thymoleptic(antidepressant and antimanic) effects of risperidone in mood disorders, this article reviews the psychotropic-induced mania, thymoleptic effects of antipsychotics, therapeutic effects of risperidone and risperidone(RIS)-induced mania(RIM) in mood disorders, risk factors of RIM, possible neurochemical mechanism of these thymoleptic effects, pathophysiological and clinical significance of thymoleptic effects, and suggestive clinical guideline of RIS in mood disorders. RIS appeared effective for bipolar disorder at a lower dose than that recommended for schizophrenia, especially in the cases of maintenance of mood stabilizers, and gradual titration from low doses. Manic induction/exacerbation can occur by chance during RIS treatment in mood disorders, schizoaffective disorders, and schizophrenias. The possible risk factors for RIM are refractory mood disorder, especially in bipolar I disorder with poor initial response ; refractory schizoaffective disorders, especially in bipolar type with poor initial response ; refractory chronic schizophrenias, especially with initial responses ; psychotic features ; higher initial doses ; rapid titration ; combined therapy with antidepressants in refractory depression ; and RIS monotherapy in mania/hypomania. RIS is a drug that preferentially block 5-HT2 receptors. The effects of low dose are due mainly to the blockade of 5-HT2 receptors. There are more gradual increase in D2 blockade with increasing dose and this D2 blocking properties become apparent at higher doses. This may be related to a modulation of dopaminergic transmission by 5-HT2 antagonism at lower doses with the direct action of RIS on DA receptors coming into play at higher dose. The serotonergic antagonistic effect may be important for its effects on depressive symptoms. This, together with adequate blo-ckade of D2 receptors, may not necessarily lead to destabilization of mood disorder, but rather to more therapeutic effects. Therefore, this dose-receptor affinity relationship with both antidepressant and antimanic effects according to treatment duration can explain a continuum of antidepressant effect, antimanic effect, behavioral stimulation, and manic/hypomanic induction/exacerbation. It was the recognition of a useful psychiatric side effects by a thoughtful observer with fertile minds that led to their ultimate utilization as psychotropic drugs, i.e., phenothiazine, MAOI, TCA, and lithium. And, in vivo pharmacological challenge by novel psychotropics, as a neurochemical probe, with more specific actions is a useful tool to select pharmacologically homogeneous subgroup of the same phenotypical(clinical) condition, to further study the unknown underlying pathogenesis of various mental illnesses. Finally, RIS may be a useful alternative or adjunctive drug for patients with mood disorders without psychotic features or refractory to treatment with standard antipsychotic drugs. The more conservative doses(tirated slowly from 1-3 mg/d) of RIS, and maintenance of mood stabilizer in the cases with risk factors of RIM are recommended in mood disorder.

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A Study on the Sediment Transport using Radioisotope Tracer (방사성동위원소 추적자를 이용한 표사이동 추적실험)

  • Choi Byung-Jong;Jung Sung-Hee;Kim Jong-Bum;Lee Jong-Sup
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.16 no.3
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    • pp.162-170
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    • 2004
  • On the basis of the radiotracer technology and the related equipments which have been developed for its industrial application through the nuclear long-term research project, a radiotracer study on sediment transport was carried out as a part of the development of the radiotracer technology for a coastal environment. The crystalline material doped with iridium having a similar composition and specific gravity as those of the bedload sand collected from the research area was produced by the oxide-route method. A radioisotope container was specially designed to inject the radiotracer from 1 m above the sea bedload without radioactive contamination during the transport from the nuclear reactor at KAERI. The position data from the DGPS and the radiation measurement data were collected concurrently and stored by means of the application software programmed with the LabVIEW of the National Instrument. The position data was reprocessed to represent the real position of the radiation probe under water and not that of the DGPS antenna on board. The time dependency of the spatial distribution of the sediment was studied in the area through three tracking measurements after the iridium glass was injected. This trial application showed the potential of the radiotracer technology as an important role for maintaining and developing the coastal environment in the future.

Genomic Organization and Characterization of the Promoter Region of Bovine ADRP (Adipocyte Different Related Protein) Gene (소 Adipocyte Differentiation Related Protein (ADRP) 유전자의 Genomic Organization 및 Promoter Region의 특성 규명)

  • Jang, Y. S.;Yoon, D. H.;Kim, T. H.;Cheong, I. C.;Jo, J. K.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.169-182
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    • 2003
  • To understand the structure and regulation of bovine ADRP (Adipocyte Differentiation Related Protein) gene, we have isolated the genomic clone of bovine ADRP and determined its sequence. A genomic Southern blot analysis confirmed that ADRP gene is present as a single copy in bovine genome and the ADRP gene spans 12 kb. Bovine ADRP genomic clone, HwADRPg-1, had 8 exons and 7 introns, and all splicing sites conformed to the GT/AG rule with the exon-intron boundaries located exactly. Analysis of the upstream 649 bp of the sequence of HwADRPg-1 showed that it does not contain any canonical TATAA boxes; however Sp1 binding sites and CAAT boxes are found. The promoter contained potential binding sites for AP-1, AP-2 and several putative transcription factor binding sites. The 5'-flanking region of HwADRPg-1 contained muscle specific transcription activator Myo G and C/EBP (CCAAT/ enhancer binding protein) recognizing site. These results suppose that the Myo G transcription activator regulate the transcription of bovine ADRP gene in muscular tissue and its transcriptional activity was triggered by degree of muscular development. Our results provide the necessary analysis for other flanking sequences are needed in addition to the proximal cis elements of this promoter to confer adipocyte differentiation-dependent or growth-dependent transcriptional control.

Molecular Cloning of Human Genomic DNA for Epinephrine Synthesizing Enzyme, Phenylethanolamine N-Methyltransferase (Epinephrine 합성효소인 phenylethanolamine N-methyltransferase의 인간 genomic DNA의 유전자 크로닝)

  • Suh, Yoo-Hun;Huh, Sung-Oh;Chun, Yang-Sook;Kim, Hun-Sik;Lim, Jung-Kyoo;Park, Chan-Woong
    • The Korean Journal of Pharmacology
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    • v.24 no.1
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    • pp.1-10
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    • 1988
  • To obtain information about the structure of the human phenylethanolamin N-methyltransferase (PNMT) and to further define the extent of the evolutionary relationships among PNMT molecules of several spesies, a full length cDNA clone for bovine adrenal PNMT was used to screen a charon 4A genomic library. One phage was isolated and identified, which included the entire PNMT gene. The length of inserted genomic DNA was 13.1-Kilobase (Kb) containing two internal EcoRI sites. Construction of a restriction map and subsequent Southern and dot blot analysis with 5'-and3'-specific cDNA probes allowed the identification of exon-containing fragments. This is the first report of the cloning of gene for human epinephrine synthesizing enzyme.

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Effects of water physico-chemical parameters on tilapia (Oreochromis niloticus) growth in earthen ponds in Teso North Sub-County, Busia County

  • Makori, Agano J.;Abuom, Paul O.;Kapiyo, Raphael;Anyona, Douglas N.;Dida, Gabriel O.
    • Fisheries and Aquatic Sciences
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    • v.20 no.11
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    • pp.30.1-30.10
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    • 2017
  • Small-scale fish farmers in developing countries are faced with challenges owing to their limited information on aquaculture management. Nile tilapia farmers in Teso North Sub-County recorded lower yields than expected in 2009 despite having been provided with required inputs. Water quality was suspected to be the key factor responsible for the low yields. This study sought to assess the effects of earthen pond water physico-chemical parameters on the growth of Nile tilapia in six earthen fish ponds under semi-intensive culture system in Teso North Sub-County. The study was longitudinal in nature with pond water and fish being the units of analysis. Systematic sampling was used to select five ponds while a control pond was purposively selected based on its previously high harvest. Four ponds were fed by surface flow and two by underground water. Each pond was fertilized and stocked with 900 fry of averagely 1.4 g and 4.4 cm. Physico-chemical parameters were measured in-situ using a multi-parameter probe. Sixty fish samples were randomly obtained from each pond fortnightly for four months using a 10 mm mesh size and measured, weighed and returned into the pond. Mean range of physico-chemical parameters were: dissolved oxygen (DO) 4.86-10.53 mg/l, temperature $24-26^{\circ}C$, pH 6.1-8.3, conductivity $35-87{\mu}S/cm$ and ammonia 0.01-0.3 mg/l. Temperature (p = 0.012) and conductivity (p = 0.0001) levels varied significantly between ponds. Overall Specific Growth Rate ranged between 1.8% (0.1692 g/day) and 3.8% (1.9 g/day). Ammonia, DO and pH in the ponds were within the optimal levels for growth of tilapia, while temperature and conductivity were below optimal levels. As temperature and DO increased, growth rate of tilapia increased. However, increase in conductivity, pH and ammonia decreased fish growth rate. Temperature and DO ranging between 27 and $30^{\circ}C$ and 5-23 mg/l, respectively, and SGR of 3.8%/day and above are recommended for higher productivity.