• Journal of Microbiology
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• v.35 no.3
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• pp.177-180
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• 1997

The purpose of this study was to evaluate the efficiency of Line Probe Assay (LiPA) in detecting the rpoB gene mutation of clinically isolated Mycobacterium tuberculosis (MTB) and to compare the level of resistance to the various rifamycins with their mutation sites. The mutation in the rpoB gene was found in 84 (97.6%) out of 86 rifampicin (RMP) resistant strains as determined by LiPA. No mutation was observed in 2 RMP resistant strains and in any of 38 RMP susceptible strains tested. Only one of 3 strains with .DELTA.5/R5, one of 2 strains with .DELTA.3, and one of 3 strains with .DELTA.2/R2 LiPA profile showed a slightly lower level of resistance to the rifapentine than the other strains. Although we could not find correlations between mutation sites in the rpoB gene and the level of susceptibility to the various rifamycins, the LiPA is recommended as a fast screening tool for detection of RMP resistant MTB.

• Journal of Microbiology and Biotechnology
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• v.7 no.6
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• pp.373-377
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• 1997

The gene for isopenicillin N synthase (cyclase; IPNS) was cloned from Lysobacter lactamgenus using DNA probe amplified with primers based on the consensus sequences of isopenicillin N synthase genes of other ${\beta}$-lactam-producing microorganisms. The genomic library of L. lactamgenus using pUC18 plasmid cloned at the SacI site were screened with the PCR-generated DNA probe and three positive clones were isolated. Enzyme activities in E. coli clones were confirmed by bioassay and HPLC assay. Throughout the functional mapping, it was observed that the gene for isopenicillin N synthase is located at the 1.3-kb XhoI-BamHI fragment of insert of positive clones. Nucleotide sequencing at both ends of the XhoI-BamHI fragment revealed that IPNS of L. lactamgenus has the common amino acid sequences at amino- and carboxy-termini.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.21 no.1
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• pp.53-66
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• 1995

The gene for ${\gamma}$-casein, a milk protein, is a member of a family of casein gene which is expressed in mammary glands of the animal during the late gestation and lactation periods binder the influence of various hormones. In order to elucidate tile mechanisms b)'which hormones regulate the coordinate induction of milk protein genes, the mouse ${\gamma}$-casein gene was isolated and characterized. The ${\gamma}$-casein gene was screened from a mouse genomic library constructed in bacteriophage EMBL3 with the ${\gamma}$-casein CDNA used as probe and one clone was obtained. The ${\gamma}$-casein CDNA as probe was partially sequenced and contained ATG start codon and 5'-noncoding region. The cloned genomic DNA was digested with Sal I restriction enzyme, by which the insert DNA can be isolated from EMBL3 vector. Three DNA bands were observed and the size of DNAs was approximately 28kb, 14kb and 9Kb, respectively Accordingly the size of the insert DNA was calculated with approximately 23Kb. The result of Southern blot analysis, however, showed that the cloned genomic DNA was not hybridized with the synthetic oligonucleotides (40 mer) of cDNA 5'-end region, but it was hybridized with the y -casein CDNA. This means that tile cloned y -casein gene may not contain its promoter region. The ${\gamma}$ -casein genomic DNA containing the promoter region has been screening from mouse genomic library with oligonucleotides of CDNA 5'-end region as probe, and twenty-nine clones was obtained.

• The Journal of Natural Sciences
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• v.5 no.1
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• pp.37-45
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• 1992

The gene coding for $\beta$-galactosidase of Neisseria lactamica 2118 was cloned into Escherichia coli MC 1061. The isolated 6.5 Kb EcoR I fragement and 7.2 Kb BamH I fragment of chromosomal DNA in Southern hybridization were ligated to a vector plasmid pBR322 and then transformed into Escherichia coli MC 1061 cells. Finally, I obtained three clones as $\beta$-galactosidase positive clone by colony hybridization and Southern hybridization($\beta$-galactosidase probe: lac Z gene of pMC1871). Three recombinant plasmids(pNL.13. 17 and 24) were found to contain the 7.2Kb BamH I fragment originated from Neisseria lactamica 2118 chromosomal DNA by Southern hybridization and pNL 24 was showed high homology to probe especially and also its physical map was constructed.

• Mycobiology
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• v.50 no.5
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• pp.382-388
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• 2022

White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.740-747
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• 2006

Amplification by universal consensus sequences in pathogenic bacterial DNA would allow rapid identification of pathogenic bacteria, and amplification of genus-specific and species-specific sequences of pathogenic bacterial DNA might be used for genotyping at the genus and species levels. For design of probes for molecular diagnostics, several tools are available as stand-alone programs or as Web application. However, since most programs can design only a few probe sets at one time, they are not suitable for large-scale and automatic probes design. Therefore, for high-throughput design of specific probes in diagnostic array development, an automated design tool is necessary. Thus, we developed a Web-based automatic system for design of genus-specific and species-specific probes for pathogen detection. The system is available at http://www.miprobe.com.

• Applied Biological Chemistry
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• v.36 no.3
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• pp.208-214
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• 1993

To investigate the gene rearrangement via short repeated sequences in chloroplast DNA, the pattern of heterologous gene clusters containing the 151 bp repeated sequence with the development of plastid was compared in rice and the homologous gene clusters from various plant sources were searched for comparative analysis. Southern blot analysis of rice DNA using rp12 gene containing 151 bp repeated sequence as a probe showed the presence of heterologous gene clusters. Such heterologous gene clusters varied with the development of plastid. Also it was observed that the heterologous gene clusters were observed in all of the rice cultivars used in this work. Finally the comparative analysis of DNA sequence of the homologous gene clusters from various plants showed the evolutionary gene rearragngement via short repeated sequence among plants. These results suggest the possible relationship between the plastid development and gene rearrangement through short repeated sequences.

• Fisheries and Aquatic Sciences
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• v.17 no.3
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• pp.339-344
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• 2014

PCR amplification with universal primer is a useful tool for speciation of symbionts in marine eukaryote coupled with robust separation method such as denaturing high performance chromatography (DHPLC). To overcome the biased amplification, clamping PCR is recommended to suppress the amplification of host gene. In this study, we evaluated the efficiency of rare gene detection for two kinds of clamping probes which were successfully utilized for eukaryotic symbiont analysis: C3 linked nucleotide (C3) and peptide nucleic acid (PNA). PNA was 3-4 orders of magnitude higher than that of C3 tested in clamping efficiency and rare gene detection. This represented that PNA could be a more competent clamping probe for the enhancement of PCR amplification for rare symbiont genes.

• Journal of Plant Biology
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• v.32 no.2
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• pp.67-78
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• 1989

Hybridization of DNA isolated from leaves of Russet Burbank potato with tomato cDNA as a probe revealed the presence of about ten inhibitor 1 genes in the genome. Screening of a genomic library of Russet Burbank potato resulted in isolation of seven different genomic clones carrying inhibitor I genes. One of the genomic clones, clone 2, contained two EcoRI fragments of 3.4 and 1.8 kb in size, respectively, which were hybridized with the probe. The nucleotide sequence of parts of the hybridizing EcoRI fragments revealed that they contain a complete gene which codes for an open reading frame of 107 amino acids. It is interrupted by two intervening sequences of 502 and 493 bp, situated at the positions of codons 17 and 43, respectively, of the open reading frame. Putative regulatory sequences, TATAAA and CCACT, were found at the 5' flanking region. In addition, a copy of a 100 bp repeat found at a tomato inhibitor I gene was identified.

• Korean Journal of Clinical Laboratory Science
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• v.42 no.3
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• pp.111-115
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• 2010

A 15-year-old female with primary amenorrhea and Tuner's syndrome feature was referred for a chromosome analysis. The karyotype of the patient was 45,X/46,X,der(Y) mosaicism under initial GTG-banding analysis. Fluorescence in situ hybridization (FISH) analysis with probe for CEP X probes and SRY probe (Vysis, Inc. Downers Grove, IL 60515, USA) was carried out. This probe is direct labeled with SpectrumOrange (SRY, Yp11.3) and is available as a single probe or mixed with the CEP X SpectrumGreen probe. SRY SpectrumOrange/CEP X SpectrumGreen hybridized to a specimen obtained from an two isodicentric Y chromosomes. The karyotype of the patient was ish Xcen(DXZ1x1)/Xcen(DXZ1x1), Yp11.3(SRYx2) by using FISH. This karyotype was considered a variant of Tuner syndrome with mixed gonadal dysgenesis (MGD), male pseudohermaphroitism (MPH) and apparently normal male.