• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.196-196
• /
• 2017

Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.

• Journal of Life Science
• /
• v.24 no.12
• /
• pp.1301-1307
• /
• 2014

Adipose-derived stem cells (ADSCs) are considered promising tools for tissue regeneration. However, ADSCs have very poor proliferation capacity. Therefore, fetal bovine serum (FBS) is generally added to the culture media of ADSCs. As FBS contains many uncharacterized components that may affect cellular functions, methods for serum-free cultures of ADSCs have been widely investigated. In this study, to develop an efficient method for a serum-free culture of ADSC-T, we used an ADSC line established by introducing the simian virus 40 (SV40) T gene into primary ADSCs. We then investigated the effect of amino acids, vitamins, and other components on the growth of ADSC-T. When the ADSC-T cells were plated with DMEM/F12 serum-free medium, the cells did not proliferate, and the mixture of amino acids, vitamins, and B27 supplement did not increase the growth of the cells. However, when the ADSC-T cells were provided with serum-free DMEM/F12 after they had been cultured with serum-supplemented DMEM for 24 h, the cells proliferated, and the vitamins and B27 supplement increased the cell growth. Stem-Pro serum-free medium also appeared to be useful as a suspension culture for the ADSC-T cells. The ADSC-T cells secreted large amounts of proteins of around 70 kDa. Insulin-like growth factor (IGF) and fibroblast growth factor basic (FGF basic) were secreted by ADSC-T in larger amounts in the serum-free culture than in the serum-supplemented culture.

• Journal of environmental and Sanitary engineering
• /
• v.18 no.1
• /
• pp.58-67
• /
• 2003

The characteristics of the commercial garbage-decomposing accelerant, Raw Material of Pomia(RMP), Pomia and Vitabio. were analyzed. In HPLC pattern, Pomia and Vitabio showed main peak of about 30kDa in 280nm wave length. RMP, Pomia and Vitabio showed three, two and two peaks in 210nm. K and Na ion content of RMP were 2,620 and 1,590ppm, respectively, and their content were the largest one among others. Ca, Mg and Mn ion content of RMP were also the largest one, but Zn ion content was the least one. $Cl^{-}$, $F^{-}$ and $NO_3^{-}$ ion content of RMP were the largest one, especially $Cl^{-}$ content was 3,553ppm. pH values of RMP and Pomia were in neutral region, but Vitabio was 9.03. Dried residue content of RMP, Pomia and Vitabio were 1.4%, 0.55% and 0.4%, respectively. The number of general bacteria on PCA plate from RMP, Pomia and Vitabio were 2.2{\times}10^{6}cfu/ml,{\;}6.5{\times}10^{3}cfu/ml{\;}and{\;}1.1{\times}10^{3}cfu/ml$, respectively. The number of facultative anaerobes of RMP was $4.4{\;}{\times}{\;}10^{4}cfu/ml$, and it was biggest one. Lactobacilli and yeasts were found less than 10cfu/ml at best. The bacterium exists in RMP in high frequency was identified as Bacillus subtilis. Volatile sulfur compound amount of garbage samples treated with Pomia and Vitabio in concentration of 1/500 at $33^{\circ}C$ for 8 days were 1,273ppb and 1,902ppb, respectively, and control showed 3,015ppb. Volatile organic compound amount of the garbage samples treated were 68,312ppb, 124,317ppb and 263,954ppm, respectively. Diethylamine that known as malodor compound were 5,107ppb, 261ppb and 11,124ppb, respectively.

• Asian-Australasian Journal of Animal Sciences
• /
• v.11 no.2
• /
• pp.121-127
• /
• 1998

Thirty two Omani sheep with eight animals each of; intact males (INT), males castrated with rubber rings immediately after birth (RR), males castrated with a burdizzo at 8 weeks of age (BC) and intact females (IF) were fed ad libitum a concentrate diet (CP 16%) plus chopped Rhodesgrass hay (8% CP) from weaning until slaughter at 28 kg. INT lambs grew faster from 9 to 20 weeks of age (p < 0.05) thus they were significantly heavier at 20 weeks of age than BC and IF, but not RR lambs. INT consumed more total feed than other sex groups over the period from 9 to 20 weeks of age. There were no significant differences between lambs of all experimental groups in feed per gain ratio. INT lambs had lower (p < 0.01) dressing percentage (DP) than RR, BC and IF. As a percentage in the empty body weight (EBW), INT had higher proportions of head, feet, empty gut (p < 0.001), liver (p < 0.05) and genitals (p < 0.05) but lower proportions of lungs and trachea (p < 0.05) than BC and IF lambs. INT males had a significantly higher (p < 0.05) proportion of bone than RR and IF lambs but lower (p < 0.05) proportion of fat than RR and IF. As a percentage in EBW, IF had the highest protortion (p < 0.001) of total body fat (TBF) followed by BC and RR whereas INT males had the lowest proportion. There was a general trend of IF having the highest proportion of individual and total non-carcass fat (TNCF) and total carcass fat (TCF) followed by BC and RR lambs whereas INT lambs had the lowest protortions of individual carcass and non-carcass fat depots. There were only few sex or castration effects on carcass tissue distribution. IF had higher proportions of intermuscular fat in the chuck, plate, leg and flank than INT and BC. The current study demonstrated that castration of intensively-raised male Omani native sheep especially at weaning using a burdizzo retarded growth rate and reduced carcass quality by increasing fat content.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.9
• /
• pp.1469-1476
• /
• 2007

Jeot-gal is a traditional Korean fermented seafood and has long been used for seasoning. We isolated 188 strains from shrimp, anchovy, and yellow corvina Jeot-gal, and screened sixteen strains that showed strong fibrinolytic activities on a fibrin plate. Among those strains, the strain that had the largest halo zone was chosen and identified as Bacillus licheniformis by using 16S rDNA sequencing and an API CHB kit. The fibrinolytic activity of Bacillus licheniformis was characterized and designated as bpKJ-31. The active component of bpKJ-31 was identified as a 37 kDa protein, designated bacillopeptidase F, by internal peptide mapping and N-terminal sequencing. The optimum activity of bpKJ-31 was shown at pH 9 and $40^{\circ}C$, with a chromogenic substrate for plasmin. It had high degrading activity for the $B{\beta}$-chain and $A{\alpha}$-chain of fibrin(ogen), and also acted on thrombin, but not skim milk and casein. The amidolytic activity of bpKJ-31 was inhibited by 1 mM phenylmethanesulfonyl fluoride, but 1 mM EDTA did not affect the enzyme activity, indicating that bpKJ-31 is an alkaline serine protease, like a plasmin. The bpKJ-31 showed approximately 14.3% higher fibrinolytic activity than the plasmin. These features of bpKJ-31 make it attractive as a health-promoting biomaterial.

• Computational Structural Engineering
• /
• v.10 no.4
• /
• pp.217-228
• /
• 1997

The high precision analysis by the p-version of the finite element method are fairly well established as highly efficient method for linear elastic problems, especially in the presence of stress singularity. It has been noted that the merits of the p-version are accuracy, modeling simplicity, robustness, and savings in user's and CPU time. However, little has been done to exploit their benefits in elasto-plastic analysis. In this paper, the p-version finite element model is proposed for the materially nonlinear analysis that is based on the incremental theory of plasticity using the constitutive equation for work-hardening materials, and the associated flow rule. To obtain the solution of nonlinear equation, the Newton-Raphson method and initial stiffness method, etc are used. Several numerical examples are tested with the help of the square plates with cutout, the thick-walled cylinder under internal pressure, and the circular plate with uniformly distributed load. Those results are compared with the theoretical solutions and the numerical solutions of ADINA

• Journal of Mushroom
• /
• v.5 no.1
• /
• pp.13-20
• /
• 2007

Analysis of genetic characteristics of Flammulina velutipes showed that strains have a range of 85% in genetic distribution diagram. According to this result, we divided these strains into five groups. In experiment of determinating the optimum media and condition in cultivating F. velutipes, we found the optimum temperature and pH range for hypha growth were $25^{\circ}C$ and 6.0 to 7.0, respectively. in addition, the best media for growth of that in plate was MCM (Mushroom Complete Media) which have a growth length from 68 to 83 mm. In vivo test, we observed that fast growth and good density of hyphae in mixture media of douglas fir sawdust, cotton seed meal and beet pulp (6:2:2 V/V). Also when we cultivated F. velutipes in this media, we harvested high yield of fruiting body.

• Journal of Microbiology
• /
• v.45 no.2
• /
• pp.158-167
• /
• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

• Journal of Life Science
• /
• v.22 no.12
• /
• pp.1658-1664
• /
• 2012

Conjugated linoleic acid (CLA) exhibits several beneficial biological activities including anticarcinogenesis and body-fat reduction. Now, we report that CLA ameliorated the oxidative stress in rat cardiomyoblast cells, H9c2, treated with hydrogen peroxide ($H_2O_2$). Cells were cultured in DMEM/F-12 media at $37^{\circ}C$ with humidified atmosphere of 5% $CO_2$. The cells, cultured for 48 hrs, were seeded at a density $3.5{\times}10^3$ cell/well in a 24 well-plate and incubated for 24 hr. Using these cells, two experiments were performed: the cytotoxicity test of CLA (10, 20, 30, 40, and $50{\mu}Ms$), and the oxidative stress amelioration test of CLA (20 and $50{\mu}Ms$) against cells treated with $H_2O_2$ (10 and 50 ${\mu}Ms$) for 1 and 2 hrs. CLA enhanced the growth of H9c2 cells at any concentrations of CLA and at any incubation times (up to 6 days), indicating that CLA acts as a growth stimulant. No protective effect of CLA (20 and $50{\mu}Ms$) was seen in cells treated $50{\mu}M$ $H_2O_2$ for 1 and 2 hr, but these CLA concentrations ameliorated (p<0.05) the adverse effect of $10{\mu}M$ $H_2O_2$ in cells treated for 1 hr. These CLA concentrations significantly (p<0.05) reduced the proportion of apoptotic cells, relative to control cells. These results suggest that CLA protected H9c2 cells from the oxidative stress of $H_2O_2$ through the suppression of cell apoptosis and could be a useful compound for the prevention of cardiac diseases caused by oxidative stress.

• Korean Journal of Food Science and Technology
• /
• v.36 no.5
• /
• pp.790-794
• /
• 2004

Bifidobacterium spp. exhibits the highest number of counts among species of microflora in breast-feeding infant intestines and has been used as probiotics. From infant groups with different diets, 42 Bifidobacterial strains were isolated by selective plate, Gram-staining, and morphology using method of Mitsuoka, among which seven isolates were identified as Bifidobacterium spp. by F6PPK test, MIDI, and PCR. B. bifidum PBH-30, selected for development of probiotics, showed high resistance against low pH and oxgall treatment, and inhibition against pathogens such as Salmonella typhimurium and Staphylococcus aureus. B. bifidum PBH-30 could be applicable to dairy products as probiotic strains due to its excellent growth in raw milk.