• Korean Journal of Microbiology
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• v.51 no.1
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• pp.31-38
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• 2015

The bacterial community structures of two marine sponges, Cinachyrella sp. and Plakortis sp., collected from Chuuk in the South Pacific in February 2012 were analyzed by PCR-DGGE (Denaturing Gradient Gel Electrophoresis) fingerprinting. After isolation of the total genomic DNAs from the sponges, the V3 regions of the 16S rRNA genes were amplified and subjected to DGGE profiling. The two species of sponges displayed different DGGE band patterns. The sequences derived from the DGGE bands revealed 85-100% similarities to known bacterial species in the public database. The bacterial community of Cinachyrella sp. was composed of 6 classes: Actinobacteria, Bacteroidetes, Chloroflexi, and Proteobacteria (Alpha-, Gamma-, Delta-). The bacterial community of Plakortis sp. included 7 classes: Actinobacteria, Chloroflexi, Firmicutes, Spirochaetes, and Proteobacteria (Alpha-, Gamma-, Delta-). Though Actinobacteria, Chloroflexi and Proteobacteria were commonly found in both sponges, the predominant bacterial communities differed between the two. Namely, the predominant bacterial groups in Cinachyrella sp. and Plakortis sp. were Proteobacteria and Chloroflexi, respectively. The sponge-associated bacteria are sponge host-specific, as each of the tested sponges from the same geographical location had different predominant bacterial diversity.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.118-122
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• 1995

Escherichia Coli ornithine transcarbamylase is the enzyme which catalyzes the L-citrulline biosynthesis from L-ornithine and carbamyl phosphate. To facilitate the purification of enzyme which will be used for many biochemical studies such as structure and function relationships and catalytic mechanisms, the cloning and expression of E. coli argI gene for ornithine transcarbamylase was conducted. argI was amplified from genomic DNA of E. coli strain of $DH5{\alpha}$, by polymerization chain reaction (PCR) method. The amplified argI gene was ligated to the prokaryotic expression vector pKK223-3 and used for transformation of E. coli TB2 which was deficient of ornithine transcarbamylase. The over-produced enzyme by the tnansformant was purified by ammonium sulfate fractionation, heat denaturation and affinity chromatography. The result of SDS denaturation gel electrophoresis for the purified enzyme showed a single band of about 38 kDa of ornithine transcarbamylase. Kinetic data for the expressed enzyme gave almost the s?????? values as those of the wild type enzyme. The $k_{cat}$, of the enzyme was $1.0{\times}10^5min^{-1}$, and $K_ms$ for ornithine and carbamyl phosphate were 0.35 mM and 0.06 mM, respectively.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.1
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• pp.26-33
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• 2006

Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.361-365
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• 1996

Indonesian Lagocephalus wheeleri ("White-milbog") and Chinese Fugu obscurus ("Hwang-bong") puffers were dissected into liver, intestine, ovary or testis, skin and muscle for assay of toxicity, Also, the toxins from the puffer liver were partially purified and analyzed. The frequency of toxic specimens for Lagocephalus wheeleri was 20, 40, 0 and 10%, in terms of liver, intestine, skin and muscle, respectively; and average toxicity was 5.5$\pm$0.9(Mean$\pm$S.E.), 11.8$\pm$4.2, 2.2$\pm$0.1 and 3.0$\pm$0.8 MU/g, respectively. All of Fugu obscurus specimens had toxicity ranging between 2 and 210 MU/g. The frequency of toxicity in intestine and ovary, both 100% was to be higher percentage tan that of other tissues. The toxinsgave two spots tetrodotoxin(TTX) and anhydro-tetrodotoxin(anh-TTX) on TLC. Also, the toxin of each puffer species showed two spots in electrophoresis. In HPLC analysis of Fugu obscurus, the toxins showed TTX, 4-epi-TTX and anh-TTX.epi-TTX and anh-TTX.

• Journal of Life Science
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• v.8 no.1
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• pp.14-26
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• 1998

a- and c-raf protein kinase in the brain of rat, the protein pattern of cerebellum during postnatal development of rat by polyacryamide gel electrophoresis, and the existence of c-raf protein kinase by using Western blotting method. The results were as follows: The cytoplasm of Purkinje cells was, in general, strongly labeled with the antibodies of a- and c-raf protein kinases in the cortex regions such as Pyramis cerebelli, Unula, Nodulus, Paraflocculus, and Flocculus. C-raf protein kinase appeared stronger immunoreactivity than a-raf protein kinase. In peripheral of cytoplasm of Nucleus emboliformis, A-raf Protein kinase was labeled markedly. During postnatal development, the protein of 38,000 dalton increased gradually in the cytosolic fraction of cerebellum, and the protein of 260,600 dalton appeared in the membrane fraction of cerebellum. By immunoblotting method, the protein band of 74,000 dalton was detected in crude and cytosolic fractions, but it was not exhibited in membrane fraction, In this fact, it was identified that a - and c-raf proteins were distributed throughout neuronal cells, especially in the Purkinje cells, in normal cerebellum cortex of rat. Also, this phenomenon was assumed that raf protein kinase in cytoplasm of neuronal cell had to do with a certain functional mechanism and signal transduction of neurotransmitter as Protein kinase C.

• Journal of Life Science
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• v.14 no.4
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• pp.702-707
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• 2004

In native-polyacrylamide gel electrophoresis of Pangasius polyuranodon, the lactate dehydrogenase (EC 1.1.1.27, LDH) $A_4$, $A_3$B, $A_2$$B_2$,$AB_3$ and $B_4$ isozymes were expressed in various tissues. The LDH $A_4$ and liver-specific $C_4$ isozymes were expressed in the tissues of Hypostomus Plecostomus. The bands of LDH in skeletal muscle, heart and eye tissues were not detected while one band was detected in kidney and liver, and four bands were detected in brain. The detected one band in liver was identified as alcohol dehydrogenase and an anodal band of skeletal muscle was identified as nothing dehydrogenase. The LDH in skeletal muscle, heart and eye might function as pyruvate reductase. The degree of inhibitions of LDH in skeletal muscle and heart of P. polyuranodon by 10 mM pyruvate were measured 57.6% and 73.8%, respectively. However, those of LDH in tissues of H. plecostomus were measured 52.7-61.8% so tissue specificity did not appear. Therefore, H. ple-costomus might be more acclimated to hypoxic environment by anaerobic metabolism of LDH iso-zymes than P. polyuranodon.

• Journal of The Korean Society of Grassland and Forage Science
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• v.25 no.4
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• pp.287-296
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• 2005

To investigation protein expression pattern in rice leaves exposed to cold stress, the soluble proteins extracted from leaf tissue were fractionated with $15\%$ PEG and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Differentially expressed proteins were identified by peptide mass fingerprinting using matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Eight proteins up-regulated and 10 down-regulated were found in $15\%$ PEG supernatant fraction. In addition, 13 proteins up-regulated and 14 down-regulated were found in $15\%$ PEG pellet fraction. It was identified the differentially expressed proteins in $15\%$ PEG supernatant fraction as pimerase/dehydratase fructokinase, ribose-5-phosphate isomerase (Rpi), chaperonin 21 precursor, probable photosystem II oxygen-envolving complex (PS II OEC) protein 2 precursor and thioredoxin h-type (Trx-h) and those in $15\%$ PEG pellet fraction as OSINBb0059K02.15, hypothetical protein, putative mitogen-activated protein kinase kinase (MAPKK), beta 7 subunit of 205 proteasome, ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit. These proteins are involved in metabolism, energy, protein synthesis, disease/defense and signal transduction-related proteins.

• Journal of The Korean Society of Grassland and Forage Science
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• v.37 no.2
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• pp.168-175
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• 2017

Single nucleotide polymorphisms (SNP) markers allow rapid screening of crop varieties in early growth stages. We developed a modified SNP PCR procedure for assaying SNPs in maize. For SNP marker development, we chosen 200 SNP sites from MaizeGDB database, and designed two base pair mismatch primers based on putative SNP site of B73 genome sequence. PCR products size was from 200 to 500 bp or was not shown in the case of SNP site existing in Korean silage corns. Using previously discovered 16 primer sets, we investigated distinctness of 50 silage F1 hybrid corns including 10 Korean silage corns developed by RDA such as Gangdaok, Kwangpyeongok, Dapyeongok, Andaok, Yanganok, Singwangok, Jangdaok, Cheongdaok, Pyeonggangok, and Pyeonganok as well as 40 foreign commercial silage corns. From cluster analysis, we confirmed that 10 Korean silage F1 hybrid corns were clearly distinguished except for Singwangok, P1395, and several foreign commercial corns, and selected minimum SNP primer combination for Gangdaok, Jangdaok, Pyeonggangok, and Pyeonganok. Therefore, development of SNP marker sets might be faster, cheaper, and feasible breed discrimination method through simple PCR and agarose gel electrophoresis.

• Journal of Animal Science and Technology
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• v.48 no.5
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• pp.719-726
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• 2006

Bovine whey protein are rich in cysteine, which is the rate limiting amino acid for synthesis of antioxidant glutathione(GSH). Some strains of Lactobacillus caseihas been reported to contain high level of GSH in cell extracts. The objective ofthis study was to determine whether enzymatically hydrolyzed whey protein isolate(WPI) and cell extract of Lb. casei HY2782 could increase intracellular GSH concentrations and protect against oxidant induced cell death in human prostate epithelial cell line (designated as RWPE1, and PC3MMM2 cells). Treatment of RWPE1 cellsandPC3MMM2 cells with hydrolyzed WPI (500g/ml) significantly increased GSH by28.2% and38.4% respectively. Compared with control cells receiving no hydrolyzed WPI(P<0.05). hydrolyzed WPI and Lb casei HY2782 cell extracts significantly protected RWPE1 and PC3MMM2 cellsfrom oxidant induced cell death compared with controls receiving no WPI. DNA damage associated with oxidant treatment was demonstrated by single cell gel (SCG) electrophoresis.

• Korean Journal of Environmental Agriculture
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• v.12 no.3
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• pp.230-238
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• 1993

To develop multifunctional microorganisms for effective wastewater treatment, the cell of P. aeruginosa P1 enable to accumulate lead in its cell were conjugated with the cell of P. fluorescens S1 enable to degrade efficiently synthetic detergents. The plasmids of the P. aeruginosa P1 and the P. fluorescens S1 were found in the cell of the conjugants when determined by agarose gel electrophoresis. The conjugants obtained from P. fluorescens S1 as a recipient cell and P. aeruginosa P1 as a donor cell possessed the ability to degrade synthetic detergents as well as to accumulate lead.