• Korean Journal of Microbiology
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• v.38 no.2
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• pp.86-95
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• 2002

As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.

• Korean Journal of Microbiology
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• v.39 no.3
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• pp.166-174
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• 2003

Microcystin produced by cyanobacteria in surface waters, such as eutrophic lake and river, is a kind of serious environmental problems due to its toxicity to human and wild animals. Microcystin is synthesized nonribosomally by the large modular multi-functional enzyme complex known as microcystin synthetase encoded by the mcy gene cluster. Amplification of mcy genes by PCR from cultures and environmental samples is a simple and efficient method to detect the toxigenic Microcystis. In order to evaluate primers designed to detect toxic microcystin-producing strains, 17 cyanobacterial strains and 20 environmental samples were examined by PCR with 7 pairs of primers. Some microcystin-producing cyanobacteria were not detected with FAA-RAA, TOX4F-TOX4R and FP-RP primers. The fragment of unexpected size was amplified with NSZW2-NSZW1 primers in Microcystis strains isolated from the lakes in Korea. TOX1P-TOX1F primers failed in amplification of toxin-producing strains. Only MSF-MSR and TOX2P- TOX2F primers amplified the fragments of mcy genes from 11 strains of microcystin-producing Microcystis. The water samples taken from 20 lakes in Korea were analyzed by PCR using each of the primers. In all the water samples, cyanobacteria capable of producing microcystin were detected by the PCR with TOX2P-TOX2F primers. These results indicate that TOX2P-TOX2F primers are better than the other primers for detection of microcystin-producing Microcystis strains in Korea. The nucleotide sequences of mcy gene in Microcystis aeruginosa NIER10010 suggest genetic diversity of Korean isolates.

• Applied Biological Chemistry
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• v.39 no.6
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• pp.443-448
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• 1996

In order to investigate the function of xylA promoter(Pxyl) as regulatory region Pxyl-lacZ fusion gene was constructed by the insertion of xylA promoter to the multiple cloning site of upstream of lacZ gene in a multicopy numbered plasmid pMC1403 containing promoterless lac operon, which was designated pMCX191, and Pxyl-lacZ fragment from pMCX191 was inserted to low copy numbered plasmid pLG339, designated pLGX191. The expressions of ${\beta}-galactosidase$ in these recombinant plasmids containing Pxyl-lacZ fusion gene were induced strongly by the addition of xylose, repressed by the addition of 0.2% glucose in the presence of xylose. The catabolite repressions were derepressed by the addition of 1 mM cAMP as same as native xylA gene. The fragment of xylA promoter was partially deleted from the upstream of xylA promoter by exonuclease III to investigate the regulation site of xylA promoter and the degrees of deletion derivatives of xylA promoter were analyzed by the DNA base sequencing. By the investigations of the induction by xylose, repression by glucose and derepression by cAMP on xylose isomerase production, the regulation site of xylA promoter may be located in segment between -165 and -59 bp upstream from the initiation site of xylA translation.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.317-323
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• 2014

Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.3
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• pp.285-290
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• 2001

To utilize canned oyster processing waste water effectively, this study was carried out to prepare instant powdered soup using the waste water (IPSW), Instant powdered souu from oyster hot-water extracts (IPSE) was prepared by mixing hot-water extracts powder (15 g) with table salt (5 g), cream powder (19 g), milk replacer (12 g), wheat flour (20 g), corn flour (15 g), starch (5 g), glucose (7.5 g) and onion powder (1.5 g). In preparing IPSW, mixed powder from wash water and boiling liquid waste, instead of powder from hot-water extracts and table salt, was added (powder from boiling liquid waste: powder from wash water= 12: 8) and other additives were added in proportion to those in the IPSE, The IPSW consists mainly of carbohydrates (about $72\%$). It was not different from the IPSE. The volatile basic nitrogen, viable cell counts, coliform group of the IPSW contains 33.4 mg/100g, $2.2\times10^4CFU/g$, <180 MPN/100g, respectively, and its water activity has 0.257. So it was a hygienically safe and conservable instant food. The main fatty acids of IPSW were 16: 0 and 18: 1n-9. Its chemical score of protein was $61.4\%$ and its main inorganic matter was iron. According to a sensual evaluation, in contrast to the IPSE, the IPSW had a bit lower aroma but better taste, It was concluded from the above chemical and sensory evaluation that even the boiling liquid waste which had been mostly abandoned because of its high table salt content can be used as a good material for instant powdered soup if it's powdered and mixed adequately with powder from wash water, and its table salt content is properly adjusted.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.166-172
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• 2002

The top shell, Omphalius pfeifferi capenteri meat vacuum-packed in can (diameter$\times$height, 74.1mm$\times$50.7mm) were heated at 115$^{\circ}C$ up to $F_0$ values of 5 min, 10 min, 15 min and 20 min, and the changes in food components were studied. After 14 days storage at 37$^{\circ}C$ and 55$^{\circ}C$, no growth of microorganism and panelling were recognized from the canned meats which were sterlized at 115$^{\circ}C$ with $F_0$ value of S min and over. In the case of proximate composition of the canned meats, the moisture content decreased with the increase of $F_0$ value, while crude protein increased. The increase of volatile basic nitrogen content, pH and degree of browning and the decrease of mineral, total amino acid, free amino acid, trimethylamine oxide, total creatinine contents and yields were observed during thermal processing, In sensory evaluation on color, texture and taste in the canned meats, no significant difference was observed among a boiled sample and the canned meats heated at re value of 10 min and below. But, in the canned meats heated at $F_0$ value of over 15 min, its sensory scores decreased with the increase of $F_0$ value. From these results, the reasonable $F_0$ value for preparation of the heat-treated top shell meats was in the range of 5$\~$10 min.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.17 no.5
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• pp.373-382
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• 1984

Processing conditions of retort pouched fried mackerel fish meat paste and quality stability during storage were investigated. The reasonable amounts of added ingredients to the frozen mackerel meat paste were $10\%$ of corn starch, $1\%$ of soybean protein, $1.5\%$ of sodium chloride, $0.6\%$ of monosodium glutamate, $0.3\%$ of alcoholic extract of red pepper, and $0.1\%$ of sodium erythorbate as an antioxidant and also added water corresponding to $10\%$ of the frozen mackerel meat paste. After grinding the defrosted mackerel fish meat paste with ingredients, the meat paste was molded in bar type and fried in soybean oil at $170-180^{\circ}C$ for 3 minutes. The fried mackerel meat paste was cooled, vacuum-packed in laminated plastic film bag (polyester/polyvinylidene chloride/unoriented polypropylene : $12{\mu}m/15{\mu}/50{\mu}m,\;14{\times}19cm$) and finally sterilized at $120^{\circ}C$ for 20 minutes in a hot water circulating retort. The pH, volatile basic nitrogen, moisture content, water activity, color, thiobarbituric acid value, peroxide value, texture and viable bacterial count of products were examined during 100 days of storage at $25{\pm}3^{\circ}C\;and\;5^{\circ}C$. The results showed that products could be preserved in good condition for 100 days at $25{\pm}3^{\circ}C$. Judging from sensory evaluation, the quality of products was not inferior to that of market products.

• Restorative Dentistry and Endodontics
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• v.30 no.5
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• pp.409-422
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• 2005

The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 XPBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing $5\%$ sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions, This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.8
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• pp.1265-1273
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• 2005

The purpose of this study was to prepare accelerated salt-fermented anchovy sauce using a shrimp processing byproducts (head, shell and tail) as a fermenting accelerator, and to investigate its physicochemical and enzymatic properties. Four types of sauces were prepared with 0, 10, 20, and 30$\%$ (w/w) addition of shrimp byproduct and fermented at 24$\pm$2$^{\circ}C$ for 360 days. During fermentation, all four type sauces decreased moisture content (67.5$\%$68.0$\%$ to 64.0$\∼$64.8$\%$) and pH (5.52$\∼$7.10 to 5.03$\∼$6.58), but showed increase in their crude protein (7.0$\∼$8.2 to 10.8$\%$) and volatile basic nitrogen contents (40$\∼$75 to 180$\∼$200 mg/100 g of sauce). The ratio of amino nitrogen to total nitrogen contents of control (0$\%$) and sauce with 10$\%$ shrimp byproducts (10$\%$ sauce) were maximized at 270 days, whereas 20$ \% $ and 30$\%$ added sauces were at 180 days. Endoprotease and exoprotease activities of anchovy sauces added with 20$\%$ and 30$\%$ of shrimp byproducts tend to be higher than those of control (0$\%$) and 10$\%$ addition. Proteolytic activities of sauces at pH 9 were about 2 times higher than those at pH 6. Amidolytic activities for LeuPNA decreased remarkably during fermentation, and control (0$\%$) almost lost their activity at 180 days, while additional sauces were relatively stable. These suggest that alkaline pretense of anchovy and shrimp byproducts as a endoprotease mainly contributed to the fermentation of salt-fermented sauces. The protein molecular weight distribution of sauces indicated 2 groups of peaks (peak 1,>70,000 da and peak 2, 3,000$\∼$29,000 da). As the fermentation proceeded, peak 1 tended to decrease in all of sauces, but peak 2 increased rapidly from 30 to 270 days. Optimum fermentation periods of control and 10$\%$ sauces were 270 days and those of 20$\%$ and 30$\%$ sauce were 180 days. The results suggest that shrimp byproduct can be used as accelerator of salt-fermented sauce.

• Journal of Korean Society of Forest Science
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• v.43 no.1
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• pp.35-38
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• 1979

The solubility variations of Abies koreana Wilson wood and their correlations ammong the factors which effect the extraction were investigated. In hot water solubility for 10 hours, extracted contents were 2.33% in heart wood, 12.30% in sapwood. In base solubility (here, used NaOH), the solubility content was 8.23% only for 1 hour. In organo-soluble fractions, there was not any variation between the content 4.00% for 1 hour and 4.44% for 10 hours, about the temperature effect, in the neutral solvent, at temperature $25{\pm}3^{\circ}C$, $50{\pm}3^{\circ}C$ and $97{\pm}3^{\circ}C$ and the solubility contents were 2.73%, 3.29%, 7.32% respectably. In the pH variations, initinal pH of solution, 6.5 became as low 5.4 after 10 hours extraction. Generally, the correlation coefficients between solubility and hour, solubility and temperature, solubilities' pH and hours, solubility and part, were r=0.890, r=0.986, r=-0.955, r=0.840 respectably. It is suggested that the most serious factor of the extraction in this material is the temperature.