• Korean Journal of Poultry Science
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• v.36 no.2
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• pp.177-186
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• 2009

It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

• Tuberculosis and Respiratory Diseases
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• v.43 no.1
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• pp.30-37
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• 1996

Background: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. Methods: We used the $InstaGene^{TM}$ DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1,245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2,536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. Results: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value. Conclusion: Even though both methods had lower possibility of cross contamination, shorter time requirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usefulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.257-264
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• 2015

To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared (FT-IR) spectroscopy can be used to discriminate and compare metabolic equivalence, standard medicinal parts from four medicinal plants (Cynanchum wilfordii Hemsley, Atractylodes japonica Koidz, Polygonum multiflorum Thunberg and Astragalus membranaceus Bunge) and their in vitro-produced adventitious roots were analyzed by FT-IR spectroscopy. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from the FT-IR spectral data showed that the whole metabolic pattern from Cynanchum wilfordii was highly similar to Astragalus membranaceus. However, Atractylodes japonica and Polygonum multiflorum showed significantly different metabolic patterns. Furthermore, adventitious roots from Cynanchum wilfordii and Astragalus membranaceus also showed similar metabolic patterns compared to their standard medicinal parts. These results clearly show that mass proliferation of adventitious roots may be applied to aquire novel supply of standard medicinal parts from medicinal plants. However, the whole metabolic pattern from adventitious roots of Atractylodes japonica and Polygonum multiflorum were not similar to their standard medicinal parts. Furthermore, FT-IR spectroscopy combined with multivariate analyses established in this study may be applied as an alternative tool to discriminate the whole metabolic equivalence from several standard medicinal parts. Thus, we suggest that these metabolic discrimination systems may be applied for metabolic standardization of herbal medicinal resources.

• Journal of Plant Biotechnology
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• v.45 no.3
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• pp.250-256
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• 2018

To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared spectroscopy (FT-IR) can be used to discriminate and compare metabolic equivalence, standard medicinal parts of Cynanchum wilfordii (Maxim.) Hemsl. and their adventitious roots were subjected to FT-IR. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from FT-IR spectral data showed that whole metabolic pattern from the adventitious root of Cynanchum wilfordii was highly similar to its standard medicinal parts. These results clearly showed that mass proliferation of adventitious roots could be applied for the novel supply of standard medicinal parts of medicinal plants. Furthermore, FT-IR spectroscopy combined with multivariate analysis established in this study could be applied as an alternative tool for discriminating of whole metabolic equivalence from standard medicinal parts. Thus, it is proposed that these metabolic discrimination systems from the adventitious root of Cynanchum wilfordii could be applied for metabolic standardization of in vitro grown Cynanchum wilfordii.

• Journal of Life Science
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• v.22 no.5
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• pp.575-581
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• 2012

This study was conducted to develop an economical optimum medium composition for the mass production of $Lactobacillus$ $plantarum$ and $Lactobacillus$ $reuteri$, livestock probiotics. Medium ingredient factors were selected on the basis of MRS broth composition, and the 15 ingredient variables were as follows: sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, dipotassium phosphate, manganese chloride, magnesium chloride, tween 80, sodium chloride, sodium acetate, ammonium citrate, sodium sulphate, and ferrous sulphate. The Plackett Burman design, consisting of 20 runs, was employed for the analysis of ingredient effects on cell growth of $L.$ $plantarum$ and $L.$ $reuteri$. As a result, sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, sodium acetate, and ammonium citrate positively influenced the growth of $L.$ $plantarum$. Additionally, yeast extract, soy peptone, $K_2PHO_4$, and tween 80 positively influenced the growth of $L.$ $reuteri$. Positive effects were found from sucrose, yeast extract, and soy peptone in the integrated analysis of the effects of both $L.$ $plantarum$ and $L.$ $reuteri$. Finally, effective medium components for both strains were found as follows: sucrose (20.0 g/l), glucose (5.0 g/l), soy peptone (11.0 g/l), yeast extract (5.0 g/l), $K_2PHO_4$ (0.2 g/l), $CH_3COONa$ (2 g/l), and $MgCl_2$ (0.02 g/l).

• Research in Plant Disease
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• v.16 no.1
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• pp.27-34
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• 2010

This study was performed to develop a formulation using an antagonistic bacterium Bacillus amyloliquefaciens A-2 to control tomato leaf mold caused by Fulvia fulva. B. amyloliquefaciens A-2 was grown in a medium with rice oil and mixed with various carrier and additives. One of the formulations, A2-MP, showed the best disease control value among the tested formulations. The disease control value of A2-MP at 100-fold and 500-fold diluted treatment was not significantly different from that of chemical fungicide triflumizole in a growth chamber. Although disease control effect was decreased by serial diluted treatment of the prepared A2-MP, 1,000-fold diluted treatment of A2-MP still showed high disease control value of 72.0%. For the green house experiments, the disease control values of A2-MP was indicated as 79.4% which is similar to that of chemical fungicide, triflumizole showing 79.6%. When the disease control activity of the formulation A2-MP was compared in tomato production conditions, disease control values of 100-fold diluted A2-MP and 3,000 fold diluted triflumizole exhibited 60%, 81.6%, respectively. The disease control efficiency by A-2MP was 73% of the disease control value of chemical fungicide. The formulation A-2MP maintained the stable bacterial viability and disease control activity when stored at $4^{\circ}C$. This result suggested that A-2MP develped from B. amyloliquefaciens A-2 could be used to control tomato leaf mold.

• Journal of Life Science
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• v.30 no.2
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• pp.156-161
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• 2020

Since industrialization, the production and utilization of various chemicals has contributed to improving the quality of our lives, but the subsequent discharge of massive waste is inevitable, and environmental pollution is becoming more serious every day. Exposure to chemicals as a result of environmental pollution is having a negative effect on human health and the ecosystem, and cleaning up the polluted environment that can affect our lives is a very important issue. Toxic aromatic compounds have been detected frequently in soil, groundwater, and wastewater because of the extensive use of oil products, and phenol, which is used to produce synthetic resins, textiles, and dyes, is one of the major pollutants, along with insecticides and preservatives. Phenol can cause dyspnea, headache, vomiting, mutation, and carcinogenesis. Phenol-degrading bacterium DWB-1-8 was isolated from the activated sludge of textile wastewater; this strain was identified as Comamonas testosteroni by 16S rRNA gene sequencing. The optimal culture conditions for the cell growth and degradation of phenol were 0.7% K₂HPO₄, 0.6% NaH₂PO₄, 0.1% NH₄NO₃, 0.015% MgSO₄·7H₂O, 0.001% FeSO₄·7H₂O, an initial pH of 7, and a temperature of 30℃. The strain was also able to grow by using other toxic compounds, such as benzene, toluene, or xylene (BTX), as the sole source of carbon.

• Journal of Aquaculture
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• v.21 no.3
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• pp.157-166
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• 2008

Species composition and standing crop of phytoplankton were investigated in the ark shell Scapharca broughtonii farming areas from March, 2006 to February, 2007 in Jinhae Bay. Water temperature ranged from 7.56 to $25.90^{\circ}C$, salinity from 13.74 to 34.78 psu, dissolved oxygen from 4.13 to 13.20 mg/L, chlorophyll $\alpha$ from 2.77 to 104.98 $mg/m^2$ and pH from 7.83 to 8.65. Dominant species was Nitzschia and Rhizosolenia from March to May, Skeletonama costatum and Prorocentrum from June to August, Skeletonama costatum, Thalassiosira, Chaetoceros from September to November and Thalassiosira, Chaetoceros from December to February. Colonial diatoms were more dominant than the single cell diatoms. Standing crop was the highest in July at three stations. Standing crop of Skeletonama costatum was the highest as 1,760.0 cells/mL at St. 1, 1,075.2 cells/mL at St. 2 and 698.4 cells/mL at St. 3 in July.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.319-328
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• 1997

To develop a beneficial microbial feed for the cultivation of rotifer, Brachionus plicatilis, an aerobic photosynthetic bacterium, Erythrobacter sp. $S\;\pi-I$ was isolated from marine structure at Haeundae beach in Pusan, Korea. Feeding effects of Erythrobacter sp. $S\;\pi-I$ on the growth of rotifer were analyzed comparing to other feeds such as PSB (purple nonsulfur bacteria), Chlorella sp. and baker's yeast. Erythrobacter sp. $S\;\pi-I$ contained more linoleic acid $(C_{18:3\omega3})$ and oleic acid $(C_{18:1\omega9})$ and amino acids than PSB (purple nonsulfur bacteria), Chlorella sp. and baker's yeast. The rotifer fed on Erythrobacter sp. $S\;\pi-I$ showed better effects than those fed on other feeds in the individual growth, size and weight. Also, the rotifer especially contained more eicosapentaenoic acid $(C_{20:5\omega3})$ and docosahexaenoic acid $(C_{22:6\omega3})$ in case of Erythrobacter sp. $S\;\pi-I$ feeding than the other feeds. In case of the feed of PSB and baker's yeast docosahexaenoic acid $(C_{22:6\omega3})$ did not show. In amino acid analysis, the rotifer fed on Erthrobacter sp, $S\;\pi-I$ showed more amino acid content comparing to those fed on other diets. Especially, arginine, isoleucine, histidine, lysine, methionine, phenylalanine, threonine, which are essential amino acid for fish growth, showed high contents. These results suggested that the aerobic photosynthetic bacterium, Erythrobacter sp. $S\;\pi-I$ would be a beneficial microbial teed for the cultivation of rotifer.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.5
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• pp.674-680
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• 2005

The purposes of this study were to compare the effect of several reheating treatments (heating in the frying pan, convection oven and microwave oven) on sensory characteristics and to evaluate the safety during storage period of cook/chill Pajeon. The sensory evaluations were made on 5 sensory attributes by a 9-member panel using quantitative descriptive analysis (QDA). The fresh cooked Pajeon and the Pajeon reheated in the frying pan obtained a significantly (p<0.01) higher score in taste than the ones reheated in a convection oven and microwave oven. The reheated cook/chill Pajeon had a significantly (p<0.01) lower score in flavor than the freshed cooked one. Regardless of the reheating methods, sensory scores in texture of the Pajeon reheated at $v$ for 1 day were not different from that of fresh cooked one. However, the scores of the reheated ones in a convection oven and in a microwave oven decreased with storage time up to 5 days at $3^{\circ}C$. On the other hand, the Pajeon reheated in the frying pan, even after 3 days' storage at $3^{\circ}C$, was not found to be inferior to the freshed cook one in every quality attributes except flavor. Therefore, the reheating treatment in frying pan may be superior to those in a convection oven and a microwave oven. The safety of Pajeon was also evaluated by measuring total count, coliform count, psychrotrophic count, acid value and peroxide value during 5 days of storage periods at $4^{\circ}C$. Total counts of Pajeon was ranged from not detectable to $5.2\times10^2$ CFU/g. The coliform and psychrotroph were not detected at all experiments. The acid values were ranged from 1.90 to 4.03 mg of KOH/g of fat until 5 days at $4^{\circ}C$. And the peroxide values were ranged from 3.63 to 12.50 meq of peroxide/kg of fat until 5 days of storage period. Therefore, these results demonstrated that Pajeon is microbiologically and chemically safe during 5 days of storage period at refligeration temperature.