• Microbiology and Biotechnology Letters
• /
• v.13 no.1
• /
• pp.87-91
• /
• 1985

Hpa I endonuclease from Haemophilus parainfluenzae has been purified of homogeneity and its physical and ezymatic properties have been studied. For the purification of the enzyme, Heparin agarose, SP-sephadex C-25, DEAE-sephadex A-50 and phosphocellulose chromatography columns were used. The denatured and reduced form of the enzyme is a monomer of molecular weight of $30,000{\pm}1,000$ as judged by 10% polyacrylamide gel electrophoresis containing 0.1% sodium dodesyl sulfate. Hpa I endonuclease was maximally active at neutral pH (7.0 to 7.5) in the presence of 50 mM NaCl.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.18 no.6
• /
• pp.557-562
• /
• 1985

Moderately halophilic bacteria were collected from solar salt with Larsen medium containing $10\%$ NaCl. A total of 56 strains were isolated and tested for the presence of plasmid DNA by agarose gel electrophoresis. Twelve isolates ($21\%$) carried at least one kind of plasmid. Six different isolates among them were selected to study the molecular weight of plasmids and the morphological and physiological characters. Vibrio sp. 14, Alcaligenes sp. 63, Pseudomonas sp. 11, Flavobacterium sp. 38, Bacillus sp. 16, and Alcaligenes sp. 52 carried at least one plasmid of about 7.2 kbp, 6.4 kbp, 6.85 kbp, 8.5 kbp, 8.75 kbp, and 6.8 kbp respectively.

• Korean Journal of Microbiology
• /
• v.40 no.4
• /
• pp.359-363
• /
• 2004

I investigated an effective way to generate a metagenomic library from DNA prepared from environental samples. The sizes of DNA extracted from environmental samples were usually in the range of 10 to 100 kbp as estimated from $0.4\%$ agarose gel electrophoresis. Because of this small size, a fosmid, rather than BAC, was chosen as a vector. It was found that, for the successful generation of metagenomic library, the selection of DNA with the sized of about 40 kbp was critical and, therefore, a simple agarose gel electrophoresis system was developed to select this size of DNA. By the procedure described in this report, I obtained metagenomic libraries containing 25,000 fosmid clones, which corresponded to 1,000 Mb of metagenomic DNA.

• Journal of Crop Science and Biotechnology
• /
• v.10 no.4
• /
• pp.237-242
• /
• 2007

Genome duplication(i.e. polyploidy) is a common phenomenon in the evolution of plants. The objective of this study was to achieve a comprehensive understanding of genome duplication for SNP discovery by Thymine/Adenine(TA) cloning for confirmation. Primer pairs were designed from 793 EST contigs expressed in the roots of a supernodulating soybean mutant and screened between 'Pureunkong' and 'Jinpumkong 2' by direct sequencing. Almost 27% of the primer sets were failed to obtain sequence data due to multiple bands on agarose gel or poor quality sequence data from a single band. TA cloning was able to identify duplicate genes and the paralogous sequences were coincident with the nonspecific peaks in direct sequencing. Our study confirmed that heterogeneous products by the co-amplification of a gene family member were the main cause of obtaining multiple bands or poor quality sequence data in direct sequencing. Counts of amplified bands on agarose gel and peaks of sequencing trace suggested that almost 27% of nonrepetitive soybean sequences were present in as many as four copies with an average of 2.33 duplications per segment. Copy numbers would be underestimated because of the presence of long intron between primer binding sites or mutation on priming site. Also, the copy numbers were not accurately estimated due to deletion or tandem duplication in the entire soybean genome.

• The Korean Journal of Zoology
• /
• v.25 no.3
• /
• pp.123-129
• /
• 1982

In order to establish a biochemical genetic system in Drosophila populations in Korea, the $\\alpha$-glycerophosphate dehydrogenase alleles of eleven natural populations of D. melanogaster in Korea were examined by means of agarose gel electrophoresis. The results obtained are presented below: 1. $\\alpha$-Glycerophosphate dehydrogenase ($\\alpha$-GPDH) allele is scored for eleven natural populations of D. melanogaster in Korea, resulting that $\\alpha$-GPDH is found to be widely polymorphic for two electrophoretic variants. 2. The heterozygosity of $\\alpha$-GPDH is calculated as $40\\sim50%$. 3. The frequency of the FF genotype of $\\alpha$-GPDH is found to be roughly same as the SS genotype, but less than the FS genotype. 4. The F gene of $\\alpha$-GPDH is distributed almost frequently as the S gene.

• Microbiology and Biotechnology Letters
• /
• v.13 no.4
• /
• pp.345-348
• /
• 1985

Recombinant chimeric plasmid constructed with Xba I digested pUBl10 and -pE194 was transformed by polyethylene glycol induced protoplast transformation system into Bacillus subtilis BR 151 on the mannitol regeneration media, and two genes of antibiotics resistance were expressed simultaneously in the transfromant. Transformation frequency of the recombinant plasmid was 6.5 $\times$ 10$^{-5}$ on the mannitol regeneration agar plate containing neomycin and erythromycin. The replication of recombinant plasmid in the recipient cells was confirmed by the alkaline extraction method and agarose gel electrophoresis.

• Environmental Mutagens and Carcinogens
• /
• v.17 no.2
• /
• pp.71-77
• /
• 1997

The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

• Proceedings of the KSME Conference
• /
• 2008.11a
• /
• pp.1411-1414
• /
• 2008

It is reported that mechanical stimulation takes a role in improving cell growth in skeletal system. And various research groups have showed that developed bioreactor to stimulate cell-seeded and threedimensional scaffold. In this study, we designed a custom-made bioreactor capable of applying controlled compression to cell-seeded agarose gel. This device consisted of a circulation system and compression system. In circular system, culture chamber was sealed for prohibiting contamination and media solution was circulated by pump. In compression system, mechanical stimuli were controlled by LabVIEW software and mechanical transfer system. Cell-encapsulated agarose gels were cultured for up to 7 days. There were significant differences between the number of cells grown in dynamic cell culture and in static cell culture from 3 days to 7 days.

• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.142.1-142.1
• /
• 2003

Sensitive and safe method for visualization of DNA in agarose gels using visible dye is described. To improve the sensitivity, we studied a counterion-dye staining method using methyl orange as a counterion-dye which contributes to reduce excessive background staining by indoine blue. Dye concentrations, PH of staining solution, mixing molar ratio of two dyes, and staining times were optimized for the counterion-dye staining. By the staining with a mixed solution of 0.005% indoine blue and 0.00165% methyl orange in 10% ethanol 0.2M sodium acetate, 8 ng of the 3 kb DNA in an agarose gel was detected within 1hr. (omitted)

• The korean journal of orthodontics
• /
• v.9 no.1
• /
• pp.9-14
• /
• 1979

The purpose of this study was to pursue the biosynthesis of proteins of human and bovine periodontal ligaments in vitro system. The excised periodontal ligaments from human and bovine were incubated in Krebs-glucose medium containing $^3H$-proline. After incubation the incubated periodontal ligaments were homogenized and the proteins were treated with 0.1%sodium dodecyl sulfate and $\beta$-mercaptoethanol. Separation of the protein fractions was performed with agarose gel column chromatography and SDS acrylamide gel electrophoresis. The results indicated as follow: 1. Only a small percentage of $^3H$-proline incorporated into proteins was hydroxylated to $^3H$-hydroxyproline. 2. The labeled proteins in periodontal ligaments showed a wide distribution of molecular weight. But only small amounts of labeled protein were found that were characteristics of the molecular weight of collagen. 3. In all of the combined fractions of gel filtration, the degree of hydroxylation was small.