• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2017년도 9th Asian Crop Science Association conference
• /
• 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.

• 생명과학회지
• /
• 제24권12호
• /
• pp.1301-1307
• /
• 2014

지방유래줄기세포(adipose-derived stem cell; ADSC)는 조직재생을 위한 탁월한 수단으로 인정되고 있으나 세포증식속도가 느려 ADSC의 배양용 배지에는 대게 fetal bovine serum (FBS)이 첨가된다. FBS는 세포에 다양한 영양분을 공급하지만 세포의 기능에 영향을 미칠 수 있는 미 동정 물질도 많이 함유하고 있다. FBS에 의한 예상밖의 영향과 동물유래물질의 오염을 방지하기 위해 ADSC의 무혈청 배양법에 관한 연구가 광범위하게 이루어지고 있다. 본 연구에서는 ADSC세포에 SV40의 T항원 유전자를 도입하여 증식속도를 향상시킨 ADSC-T세포주의 효율적인 무혈청 배양법을 확립하기 위해 ADSC-T의 세포증식에 미치는 아미노산복합체, 비타민 복합체 및 여러가지 영양분 혼합물(B27)의 영향을 검토하였다. 그 결과, ADSC-T세포를 DMEM/F12 무혈청 배지에 현탁하여 plate에 주입하였을 때는 증식하지 않았으며 아미노산, 비타민 및 B27 영양소복합체는 증식촉진효과를 나타내지 않았다. 그러나 ADSC-T세포를 유혈청 DMEM배지로 24시간 배양 후 DMEM/F12 무혈청 배지로 교체하여 배양했을 때는 세포가 증식하였으며 이때, 비타민 복합체와 B27 영양소복합체는 증식촉진효과를 나타내었다. 또한 Stem pro 배지를 이용하면 ADSC-T의 무혈청 부유배양이 가능한 것으로 나타났다. ADSC-T세포는 분자량 70 kDa 부근의 단백질을 다량으로 분비하였으며, 성장인자 중에서 insulin-like growth factor (IGF)와 fibroblast growth factor basic (FGF basic)는 유혈청 배양보다 무혈청 배양에서 더 많이 분비되었다.

• 환경위생공학
• /
• 제18권1호
• /
• 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
• /
• 제11권2호
• /
• 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
• /
• 제17권9호
• /
• 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.

• 전산구조공학
• /
• 제10권4호
• /
• pp.217-228
• /
• 1997

p-version 유한요소법에 의한 고정밀해석은 응력특이가 발생하는 선형탄성 문제에 매우 적합한 방법으로 인식되고 있다. 해석 결과의 정확도, 모델링의 단순성, 입력자료에 대한 통용성 및 사용자와 CPU 시간의 절감 등 여러장점이 선형탄성 문제에 적용되어 우수성이 입증되었지만, 탄소성 해석분야는 아직 적용이 이루어지지 않고 있다. 그러므로 본 논문에서는 일-경화재료에 대한 구성방정식을 이용하여 정식화된 증분소성이론과 소성유동법칙에 근거한 재료비선형 p-version 유한요소모델이 제안되었다. 비선형방정식을 풀기 위해 Newton-Raphson법과 초기강성도법 등의 반복법이 모색되었다. 제안된 모델을 이용하여 개구부를 가진 사각형 평판과 내압을 받는 두꺼운 실린더, 그리고 등분포하중을 받는 원판해석 등의 수치실험이 수행되었다. 한편, p-version 모델에 의한 해석결과는 문헌의 이론값과 상용유한요소프로그램인 ADINA의 해석결과와 비교 검증되었다.

• 한국버섯학회지
• /
• 제5권1호
• /
• pp.13-20
• /
• 2007

갈색 팽이균주간의 유전적 특성을 보기 위하여 RAPD를 수행하여 유전적 계통도를 본 결과 85% 수준에서 5개의 그룹으로 구분되었다. 생리적 특성으로는 최적 온도는 $25^{\circ}C$이고, CBFM-002를 제외한 모든 균주가 pH 6.0~7.0 범위에서 균사 생장이 양호하였다. 균사생장은 MCM 배지에서 68.4~83.4 mm로 가장 좋았고 균사밀도는 WA배지를 제외한 모든 배지에서 균사밀도가 높았다. 균총과 자실체 간의 색깔 간에는 일정한 경향을 찾을 수 없었다. 갈색팽이버섯 재배에 적당한 배지 재료를 선발하기 위하여 검토한 결과, 배지별 이화학적 성분 분석에서 T-N은 미송+면실박+비트펄프 (6:2:2)가 가장 많았고, $P_2O_2$, $K_2O$, MgO는 미송+쌀겨(8:2)배지에서 가장 많이 함유되어 있었다. 배지혼용 처리에 따른 균사생장 정도는 CBFM-003, CBFM-007균주 모두 미송+면실박+비트펄프 (6:2:2) 배지에서 균사생장이 빠르고 균사밀도가 높았다. 혼용된 배지로 병재배시 자실체의 갓 직경, 대 직경, 대 길이는 CBFM-003, CBFM-007 균주는 모든 배지에서 비슷한 경향을 보였다. 병버섯 재배에서 배지 혼합비율에 따른 균주간 균사 및 자실체의 생육은 미송+쌀겨 (8:2)보다 미송+면실박+비트펄프 (6:2:2)배지에서 CBFM-003은 156.3 g, CBFM-007은 162.8 g으로 가장 좋은 수량을 보였다. 병재배에서 생육온도에 따른 수량은 $7^{\circ}C$에서 약간 높은 수량을 보였으나, 발이기간 동안 에너지 투입량 등 경제성을 고려하면 CBFM-007은 발이온도 $10^{\circ}C$가 적합한 것으로 판단되었다.

• Journal of Microbiology
• /
• 제45권2호
• /
• 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.

• 생명과학회지
• /
• 제22권12호
• /
• pp.1658-1664
• /
• 2012

Hydrogen peroxide ($H_2O_2$)를 처리한 rat cardiomyoblast H9c2 cell (rat 배아심근 세포)에서 conjugated linoleic acid (CLA)의 oxidative stress 경감 효과를 연구하였다. H9c2 세포는 DMEM/F-12 배지에서 배양($37^{\circ}C$, 5% $CO_2$)하였다. CLA (10, 20, 30, 40, $50{\mu}M$)는 배양 6일간 H9c2 세포 증식을 거의 직선적으로 촉진하였다. 따라서 CLA ($20{\mu}M$, $50{\mu}M$)를 H9c2 cell에 2일간 처리한 후 $H_2O_2$ ($10{\mu}M$; 각각 1시간 및 2시간 처리)의 독성을 조사한 결과 CLA ($20{\mu}M$, $50{\mu}M$)는 $10{\mu}M$ $H_2O_2$ 1시간 처리에서 세포독성을 대조에 비해 유의성 있게 억제하였다(p<0.05). 또한 이 CLA는 apoptotic DNA 분포를 대조 세포에 비해 유의성 있게 감소 시켰다(p<0.05). 따라서 이 결과는 CLA가 $H_2O_2$에 의한 apoptosis를 억제함으로서 oxidative stress로부터 H9c2 세포를 보호한다고 사료되고, 이 CLA는 cardiac diseases를 보호할 수 있는 물질로 사용될 수 있을 것이다. 그러나 많은 연구가 수행되어야 할 것이다.

• 한국식품과학회지
• /
• 제36권5호
• /
• pp.790-794
• /
• 2004

본 연구에서는 유아용 생균제 개발을 위하여 1세 미만의 한국인 유아로부터 Bifidobacterium균을 분리 및 동정하여 생균제 특성이 우수한 균주를 선발하고자 하였다. 20명의 충북지역 유아 분변으로부터 선택배지를 이용한 plate법, 그람염색법을 이용하여 42종의 Bifidobacterium균을 분리하였고, F6PPK test, MIDI 및 PCR 방법을 통해 최종 7종의 Bifidobacterium균을 동정하였다. 동정된 7종의 Bifidobacterium균의 내산성 및 내담즙성 실험을 통하여 높은 효능을 나타내는 B. psedocatenulatum PBH-29와 B. bifidum PBH-30을 선발하였다. 그리고 병원성균 성장억제 실험에서 B. bifidum PBH-30이 Sal. typhimurium과 Staph. aureus에 대하여 B. psedocatenulatum PBH-29보다 효과적인 저해를 유도하였고, 10% skim milk에서도 48시간 동안 배양한 결과, B. bifidum PBH-30이 B. psedocatenulatum PBH-29보다 좀 더 높은 성장능을 나타냈다. 따라서 본 연구에서는 최종적으로 B. bifidum PBH-30을 유아용 생균제 개발을 위한 균주로 선발하였다.