• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.2
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• pp.128-133
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• 1998

Analyses of now karyotypes using different maize (Zea mays L.) inbred lines have been performed. The accumulation and isolation of high quality and quantity metaphase chromosomes from root tips can be achieved from many kinds of maize lines. The chromosome suspensions were prepared by a simple slicing method from synchronized maize root tips and analyzed with a now cytometry. The variations of experimental now karyotypes were detected among inbred lines in terms of the positions and/or the numbers of chromosome peaks. The 2C DNA amount among 8 inbred lines ranged from 5.09 to 5.52 pg. The variability of DNA content in maize chromosome 1 was 9.1 % ranging from 0.685 to 0.747 pg. The selection of appropriate maize lines is critical for sorting specific single chromosome types. At least five different chromosome types can be discriminated and sorted from five maize lines.

• Genomics & Informatics
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• v.4 no.2
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• pp.80-86
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• 2006

It is clear that the construction of large insert DNA libraries is important for map-based gene cloning, the assembly of physical maps, and simple screening for specific genomic sequences. The bacterial artificial chromosome (BAC) system is likely to be an important tool for map-based cloning of genes since BAC libraries can be constructed simply and analyzed more efficiently than yeast artificial chromosome (YAC) libraries. BACs have significantly expanded the size of fragments from eukaryotic genomes that can be cloned in Escherichia coli as plasmid molecules. To facilitate the isolation of molecular-biologically important genes in Ashbya gossypii, we constructed Ashbya chromosome-specific BAC libraries using pBeloBAC11 and pBACwich vectors with an average insert size of 100 kb, which is equivalent to 19.8X genomic coverage. pBACwich was developed to streamline map-based cloning by providing a tool to integrate large DNA fragments into specific sites in chromosomes. These chromosome-specific libraries have provided a useful tool for the further characterization of the Ashbya genome including positional cloning and genome sequencing.

• The Korean Journal of Zoology
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• v.29 no.3
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• pp.208-214
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• 1986

Comparative study of karyotypes in two allotypes $(Mdh-1^{MM} and Mdh-1^{MS})$ of the dark chub (Zacco temmincki) was examined. Both types had diploid number of 48 but the 7th chromosome was strikingly different between them. The chromosomes of $Mdh-1^{MM}$ type was consisted of 6 pairs of metacentrics, 6 pairs of submetacentrics, and 12 pairs of acrocentrics whereas the chromosomes of $Mdh-1^{MS}$ type had 7 pairs of metacentrics, 5 pairs of submetacentrics and 12 pairs of acrocentrics. No hybrid type between these two types was found in sympatric area at Tongchon River Namhae. Probable reproductive isolation between them was discussed.

• Korean Journal of Breeding Science
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• v.40 no.3
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• pp.234-242
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• 2008

To identify low temperature-induced genes of wheat chromosome substitution line 5D, suppression subtractive hybridization (SSH) was performed with mRNAs from leaf samples that treated with low temperature ($4^{\circ}C$). A cDNA library was constructed using mRNA isolated from wheat chromosome substitution line 5D leaves treated with low temperature ($4^{\circ}C$). The nucleotide and deduced amino acid sequences of the putative gene products were compared. wfr-9 and wfr-32 showed identity over 90% related to vernalization gene. Other two genes, wfr-77 and wfr-83 which is related to freezing-resistant gene have also identity over 90%. This result suggest that those genes may be transcribed into antifreeze proteins which are accumulated within leaf apoplasts, when wheat chromosome substitution line 5D is acclimated during low temperature treatment.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1079-1082
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• 2007

Chromosome microdissection and the reverse FISH technique is one of the most useful methods for the identification of structurally abnormal chromosomes. In particular, the laser microbeam microdissection (LMM) method allows rapid isolation of a target chromosome or a specific region of chromosomes without damage of genetic materials and contamination. Isolated chromosomes were directly amplified by the degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), and then the FISH probes labeled with spectrum green- or spectrum red-dUTP were generated by nick-translation. Whole chromosome painting (WCP) probes were successfully generated from only 5 copies of the chromosome. With this method, we produced 24 WCP probes for each human chromosome. We also tried to characterize a marker chromosome, which seemed to be originated from chromosome 11 on conventional banding technique. The marker chromosomes were isolated by the LMM method and analyzed by reverse FISH. We elucidated that the marker chromosome was originated from the short arm of chromosome 5 ($5p11{\to}pter$). A fully automated and computer-controlled LMM method is a very simple laboratory procedure, and enables rapid and precise characterization of various chromosome abnormalities.

• Korean Journal of Veterinary Service
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• v.33 no.3
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• pp.233-240
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• 2010

Methicillin-resistant coagulase-negative staphylococci (MRCNS) were isolated from the respiratory sites of chickens in 4 farms and slaughter house located in Chungnam provinces. Isolation of coagulase-negative staphylococci (CNS) was positive for 61 (26.6%) of the 229 chickens tested, and isolation of MRCNS was positive for 17 (27.9%) of the isolated CNS. A total of 17 MRCNS isolates were selected and subjected to identification. Of the 17 MRCNS isolates selected, 6 were identified as Staphylococcus cohnii, 2 as S. saprophyticus, 3 as S. simulans, 3 as S. lentus, 2 as S. carnosus, and 1 as S. xylosus. The MRCNS isolates were resistant to many beta-lactam antibiotics, and some isolates were also resistant to macrolide and aminoglycoside antibiotics. The mecA gene was detected in some isolates of each MRCNS strains. The mecA-positive isolates were classified into five staphylococcal cassette chromosome mec (SCCmec). SCCmec types I to IV were detected in isolates from chickens.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.388-398
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• 1998

A bacterium which is resistant to both mercury and cadmium, and also capable of utilizing phenol as a carbon and energy source, was isolated from the Kumho River sediments near Kangchang Bridge, Taegu, Korea. The isolate was labeled Pseudomonas sp. KM10 and characterized. The bacteria grew in 4 mM $CdCl_2$and in $70{\mu}M$ $HgCl_2$. The bacteria efficiently removed over 90% of 1 g/l phenol within 30 h. In the presence of 1.250 g/l phenol, the growth of the microorganism was slightly retarded and the microorganism could not tolerate 1.5 g/l phenol. Curing of plasmid from the bacteria was carried out to generate a plasmidless strain. Subsequent experiments localized the genes for phenol degradation in plasmid and the genes for mercury resistance and cadmium resistance on the chromosome. Dot hybridization and Southern hybridization under low stringent conditions were performed to identify the DNA homology. These results showed significant homologies between the some sequence of the chromosome of Pseudomonas sp. KM10 and merR of Shigella flexneri R 100, and between the some sequence of the chromosome of Pseudomonas sp. KM10 and cadA of Staphylococcus aureus pI258. The mechanism of cadmium resistance was efflux, similar to that of S. aureus pI258 cadA, and the mechanism of mercury resistance was volatilization, similar to that of S. flexneri R100 mer.

• Archives of Pharmacal Research
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• v.11 no.3
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• pp.218-224
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• 1988

The preparation of Clostridium butyricum is used as a normalizing agent for human intestinal flora. When the microbe is simultaneously used with rifampicin, it is inactivated by the antibiotic. To develop rifampicin-resistant mutants, rifampicin-sensitive strain Miyairi II 588 of C. butyricum was treated with nitrosoguanidine (NTG). To ensure stable resistance to rifampicin, we examined whether the resistance was plasmid-mediated or chromosome-mediated. It was found that the resistance of four mutant strains was not mediated by its inherent plasmid, but by the chromosomal mutation. These strains were examined for the susceptibility and resistance to other antituberculosis agents and antibiotics. The results showed that these mutants were resistant to the high concentration of the antituberculosis agents.

• Journal of Life Science
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• v.9 no.1
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• pp.69-75
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• 1999

Four species belonging to the Drosophila melanogaster complex were examined genetically and morphologically to analyze interspecific relationships. Insemination rates ranged from 96% to 99% within species crosses, but interspecific crosses among the four species exhibited a great variations in the frequency of successful matings. D. melanogaster females mated relatively well with males of other species and D. sechellia males were more successful in mating with females of other species. In the crosses among D. simulans, D. mauritiana and D. sechellia, hybrid flies were fertile in females, but sterile in males regardless of reciprocal matings. The phenogenetically relationship between this complex and their hybrids were investigated by the comparison of sex comb tooth number and genital arch of male. They were controlled by polygenic factors on the chromosome of both parents. The effects of temperature on viability of hybrids between D. melanogaster females and D. simulans males were investigated for detection of genes concerning the speciation. The temperature sensitivity of the hybrid was mainly controlled by genes located on the X chromosome of D. simulans males.

• Korean Journal of Microbiology
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• v.39 no.3
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• pp.128-134
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• 2003

Transformation-associated recombination (TAR) cloning is based on co-penetration into yeast spheroplasts of genomic DNA along with TAR vector DNA that contains 5'- and 3'-sequences (hooks) specific for a gene of interest, followed by recombination between the vector and the human genomic DNA to establish a circular YAC. Typically, the frequency of recombinant insert capture is 0.01-1% for single-copy genes by TAR cloning. To further refine the TAR cloning technology, we determined the effect of GC content on target hooks required for gene isolation utilizing the $Tg\cdot\AC$ mouse transgene as the targeted region. For this purpose, a set of vectors containing a B1 repeated hook and Tg AC-specific hooks of variable GC content (from 18 to 45%) was constructed and checked for efficiency of transgene isolation by radial TAR cloning. Efficiency of cloning decreased approximately 2-fold when the TAR vector contained a hook with a GC content ～${\leq}23$% versus ～40%. Thus, the optimal GC content of hook sequences required for gene isolation by TAR is approximately 40%. We also analyzed how the distribution of high GC content (65%) within the hook affects gene capture, but no dramatic differences for gene capturing were observed.