• 제목/요약/키워드: Et$_3$B

검색결과 782건 처리시간 0.035초

Cloning, Expression, and Characterization of a Family B-Type DNA Polymerase from the Hyperthermophilic Crenarchaeon Pyrobaculum arsenaticum and Its Application to PCR

  • SHIN HEA-JIN;LEE SUNG-KYOUNG;CHOI JEONG JIN;KOH SUK-HOON;LEE JUNG-HYUN;KIM SANG-JIN;KWON SUK-TAE
    • Journal of Microbiology and Biotechnology
    • /
    • 제15권6호
    • /
    • pp.1359-1367
    • /
    • 2005
  • The gene encoding Pyrobaculum arsenaticum DNA polymerase (Par DNA polymerase) was cloned and sequenced. The gene consists of 2,361 bp coding for a protein with 786 amino acid residues. The deduced amino acid sequence of Par DNA polymerase showed a high similarity to archaeal family B-type DNA polymerases (Group I), and contained all of the motifs conserved in the family B-type DNA polymerases for $3'{\rightarrow}5'$ exonuclease and polymerase activities. The Par DNA polymerase gene was expressed under the control of the T7lac promoter on the expression vector pET-22b(+) in Escherichia coli BL21-CodonPlus(DE3)-RP. The expressed enzyme was purified by heat treatment, and Cibacron blue 3GA and $Hirap^{TM}$ Heparin HP column chromatographies. The optimum pH of the purified enzyme was 7.5. The enzyme activity was activated by divalent cations, and was inhibited by EDTA and monovalent cations. The half-life of the enzyme at $95^{\circ}C$ was 6 h. Par DNA polymerase possessed associated $3'{\rightarrow}5'$ proofreading exonuclease activity, which is consistent with its deduced amino acid sequence. PCR experiment with Par DNA polymerase showed an amplified product, indicating that this enzyme might be useful in DNA amplification and PCR-based applications.

소, 돼지 미성숙 난포란의 유리화 동결 . 융해후 FDA 처리가 체외수정과 배 발육에 미치는 영향 (Effects of FDA Treatment after Vitrified Freezing on In Vitro Fertilization and Development of Follicular Oocytes(Bovine, Porcine) I. Survival of Mammal Follicular Oocytes after Vitrification by FDA-test)

  • 김종계;양병철;강민수;고경래;고혁진;장덕지
    • 한국수정란이식학회지
    • /
    • 제10권3호
    • /
    • pp.183-191
    • /
    • 1995
  • This experiment was carried out to study the determination of survival of vitrified and thawed mammal follicular oocytes by FDA-test. Oocytes were divided into 3 groups according to attachment of cumulus cell. Group A oocytes were tightly surrounded by cumulus cell, group B oocytes were partially surrounded by cumulus cell, and group C oocytes were poorly surrounded by cumulus cell. Vitrification solution developed by our previous study (Kim et al, 1992) which consisted of permeable agent (20 % glycerol + 10 % ethylene glycol) and nonpermeable agent (30 % Ficoll + 10 % sucrose). Oocytes (7~10) loaded into 0.25 ml straw after 10 min equilibration were plunged into liquid nitrogen (- 196$^{\circ}C$) directly. The FDA-score of vitrified and thawed group A oocytes was higher in rat (4.2) than in rabbit (3.9), cow (3.8), mouse (3.4) and porcine (2.4), however that of cumulus cell was higher in rabbit (4.7) than in rat (4.1), cow (2.9), porcine (2.6) and mouse (1.4). The FDA-score of vitrified and thawed group B oocytes were 3.1 (cow), 2.9 (rabbit), 2.9 (mouse), 2.6 (rat) and 2.5 (porcine), respectively. However that of cumulus cell was higher in rabbit (3.7) than in porcine (2.6), rat (2.3), cow (1.7) and mouse (0.3). The FDA-score of vitrified and thawed group C oocytes was higher in mouse (4.1) than in cow (2.9), rabbit (2.6), rat (1.3) and porcine (1.1). As shown in the above results, The survival rates of oocytes were higher in group A than in group B and C except in mouse and cow. These results suggest that the survival of cumulus cell as well as follicular oocytes can be reliably judged by their fluorescence with FDA-test.

  • PDF

춘천지방의 거미상 (The Spider Fauna of Chun Cheon area, Kangweon do, Korea)

  • 남궁준;백남극
    • 한국응용곤충학회지
    • /
    • 제12권3호
    • /
    • pp.131-137
    • /
    • 1973
  • This paper is given for the first time as a list of spiders from Chuncheon area, in land of kang-weon-do, mid-eastern part of Korea. Materials were collected during the early August 1972 by the Biological members of the chuncheon girl's high school, in chuncheon area. In addition to this materials collected by paik and Nam kung during July 1973 at Mt. Sam-ak which is located on south-west of chuncheon were examined. 1) In this paper, tile authors record 105 species of spiders belonging to 22 families, including 4 undetermined species from chuncheon area. 2) The followings are newly added to the spider fauna in Korea. a) Curstulina sp. b) Zelotes asiaticus(BOES. et STR.) 3) These materials are not sufficient for general discussion of the spider fauna of chuncheon area, however, an outline can be summarized as follow: 21 northern species $(20.8\%)$ 5 southern species $(5.0\%)$ 3 cosmopolitan species $(3.0\%)$ 72 plaearctic species $(71.2\%)$

  • PDF

벼 기계이앙 재배에서 벼와 잡초 경합에 따른 벼 수량 및 요방제수준 예측 (Prediction of Rice Yield and Economic Thresholds by Some Weeds-Rice Competition in Transplanted Rice Cultivation)

  • 문병철;원종건;김영림;김성우;이인용;박재읍;김도순
    • 한국잡초학회지
    • /
    • 제31권3호
    • /
    • pp.289-293
    • /
    • 2011
  • 본 연구는 벼에 대한 피, 자귀풀과 미국가막사리의 경합에 따른 수량피해 예측과 경제적인 잡초관리를 위한 방제필요 밀도를 구명하고자 하였다. 수원과 대구지역에서 얻어진 성적을 종합한 예측모델식에 따르면 논에서 피, 자귀풀, 미국가막사리가 완전히 방제되었을 때의 쌀수량은 각각 5.5t, 5.5t, 5.4t 으로 예측되었다. 초종별 평방미터당 요방제 필요밀도는 자귀풀은 제초제 종류별로 0.5~0.7본, 피는 1.2~1.4본, 미국가막사리는 1.6~1.9본인 것으로 예측되었다.

A Novel Approach to Cloning and Expression of Human Thymidylate Synthase

  • Lv, Ying-Tao;Du, Pei-Juan;Wang, Qiao-Yan;Tan, Yuan;Sun, Zong-Bin;Su, Zhong-Liang;Kang, Cong-Min
    • Asian Pacific Journal of Cancer Prevention
    • /
    • 제14권12호
    • /
    • pp.7523-7527
    • /
    • 2013
  • Thymidylate synthase (TS) catalyzes the transfer of a methyl group from methylenetetrahydrofolate to dUMP to form dTMP. It is a primary target in the chemotherapy of colorectal cancers and some other neoplasms. In order to obtain pure protein for analysis of structure and biological function, an expression vector TS-pET28b (+) was constructed by inserting wild-type human thymidylate synthase (hTS) cDNA into pET28b (+). Then an expression strain was selected after transformation of the recombined plasmid into Rosetta (DE3). Fusion protein with His-tag was efficiently expressed in the form of inclusion bodies after IPTG induction and the content was approximately 40.0% of total bacteria proteins after optimizing expression conditions. When inclusion bodies were washed, dissolved and purified by Ni-NTA under denatured conditions, the purity was up to 90%. On SDS-PAGE and West-blotting, the protein band was found to match well with the predicted relative molecular mass-36kDa. Bioactivity was 0.1 U/mg. The results indicated that high-level expression of wild-type hTS cDNA can be achieved in prokaryotes with our novel method, facilitating research into related chemotherapy.

Antifungal activity of pinosylvin from Pinus densiflora on turfgrass fungal diseases

  • Lee, Dong Gu;Lee, Seong Jun;Rodriguez, Joyce P.;Kim, Ik Hwi;Chang, Taehyun;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
    • /
    • 제60권3호
    • /
    • pp.213-218
    • /
    • 2017
  • The objective was to examine the antifungal activity of Pinus densiflora extract for the control of turfgrass fungal diseases. Antifungal activities of the various fractions of n-hexane, methylene chloride (Ch), ethyl acetate (EtOAc), and n-butanol from P. densiflora were evaluated against Rhizoctonia solani AG1-1B, R. solani AG2-2IV, Sclerotinia homoeocarpa, R. cerealis, Pythium spp., and Colletotrichum graminicola. The Ch and EtOAc fractions showed antifungal activity against Pythium sp. and C. graminicola in paper disc assay. The effective concentration to produce 50% mycelial inhibition ($EC_{50}$) using five discriminatory concentrations of pinosylvin (1) from the Ch fraction of P. densiflora was evaluated on R. solani AG1-1B, R. solani AG2-2IV, R. cerealis, and S. homoeocarpa. S. homoeocarpa showed the highest sensitivity with the lowest mean $EC_{50}$ value ($8.426{\mu}g/mL$) among the four pathogens. Among the three Rhizoctonia pathogens, R. cerealis had the highest mean $EC_{50}$ value ($99.832{\mu}g/mL$) and R. solani AG2-2IV, with the lowest sensitivity, had the lowest $EC_{50}$ value ($39.696{\mu}g/mL$). These results suggested that pinosylvin (1) from P. densiflora could be a valuable lead compound in the improvement of a novel antifungal agent.

개 외이도에서 분리한 세균 및 M pachydermatis의 약제감수성에 대하여 (Drug susceptibility of bacteria and M pachydermatis isolated from canine external ear canals)

  • 김기향;최원필;여상건
    • 대한수의학회지
    • /
    • 제39권3호
    • /
    • pp.560-565
    • /
    • 1999
  • The present work was conducted to investigate the drug susceptibility of microorganisms isolated from canine external ear canals. Antifungal susceptibility test of M pachydermatis (17 strains) was perfomed by agar dilution method, using 11 antifungal drugs including amphotericin B(A), nystatin(N), pimaricin(P), griseofulvin(G), bifonazole(B), clotrimazole(C), miconazole(M), econazole(E), ketoconazole(K), tolnaftate(T), 5-fluorocytosine(F). All isolates were highly sensitive to K, M, T(geometric mean MIC ; GM $MIC{\leq}0.16{\mu}g/ml$) but they weren't sensitive to P, F and G(GM $MIC{\geq}92.37{\mu}g/ml{\sim}{\geq}128{\mu}g/ml$). Antibacterial susceptibility test against 119 isolates of bacteria was performed by agar dilution method, using 9 antibacterial drugs including erythromycin(ET), chloramphenicol(CP), gentamycin(G), vancomycin(V), ampicillin(AP), amoxacillin(AX), chlortetracycline(CT), ciprofloxacin(CF), enrofloxacin(EF). All isolates of Staphylococcus spp(101 strains) were highly sensitive to EF, CF, G(GM MIC $0.33{\sim}1.47{\mu}g/ml$). In other gram positive cocci(4 strains), they were highly sensitive to EF, CF, V(GM MIC $1{\sim}4.76{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). In gram positive rods(13 strains), they were highly sensitive to EF, CF, G(GM $MIC{\leq}0.19{\sim}1{\mu}g/ml$). In Pseudomonas aeruginosa(1 strain), it was highly sensitive to AX, EF, ET, CF(GM MIC $0.06{\sim}1{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). All isolates weren't sensitive to AP(GM MIC 16~>$32{\mu}g/ml$).

  • PDF

파형벽면이 있는 채널내의 난류열전달에 대한 수치해석 (Numerical Simulation of Turbulent Heat Transfer in a Channel with One Wavy Wall)

  • 박태선
    • 한국추진공학회지
    • /
    • 제9권3호
    • /
    • pp.49-59
    • /
    • 2005
  • 파형벽면이 존재하는 채널유동에 대한 난류열전달이 난류모델에 의해서 조사되었다. Park et at.[1]의 비선형 k- f - f$_{모델이 수정되었고, 외재적인 비선형 열유속모델이 사용되었다 선택된 레이놀즈수는 Re$_{b}$ =6760이고 형상변화 (0 $\leq$ $\alpha$/$\lambda$$\leq$0.15 and 0.25 $\leq$A/H$\leq$4.0.)에 따른 열전달을 조사하였다. 모델의 성능을 검증하기 위하여 큰에디모사법이 선택된 경우에 수행되었다. 큰에디모사법의 결과와 비교할 때 모델성능은 일반적인 경향을 잘 예측하였다. 비선형 k- $\varepsilon$ - f$_{모델을 이용하여 파형벽면에 의한 열전달의 증가 특징과 형상의 영향이 조사되었다.

서해안 저소성 점토질 실트 지반의 부분배수 특성 (Partial Drainage Characteristics of Clayey Silt with Low Plasticity from the West Coast)

  • 김석조;이상덕;김주현
    • 한국지반공학회논문집
    • /
    • 제32권9호
    • /
    • pp.17-27
    • /
    • 2016
  • 서해안의 인천 및 화성지역에 분포하는 모래 및 실트 함유량이 많은 저소성 지반에 대해 피에조콘관입시험(CPTU) 데이터 및 강제치환 공법을 이용하여 부분배수 특성을 분석하였다. Powell과 Quarterman(1988)에 의한 과압밀비 $OCR={\kappa}(q_t-{\sigma}_{vo})/{\sigma}^{\prime}_{vo})$ 경험식은 모래함유량이 많은 서해안 저소성 실트 지반에서는 상대적으로 투수성이 커서 표준관입속도(2cm/s)하에서 콘관입저항력($q_t$)이 크게 평가되어 과압밀비가 크게 산정되는 경향을 나타냈다. Schnaid et al. (2004)는 간극수압계수($B_q$)-강도증가율($s_u/{\sigma}^{\prime}_{vo}$)-정규화된 콘저항($Q_t=(q_t-{\sigma}_{vo})/{\sigma}^{\prime}_{vo}$)을 함께 도시하여, 부분배수 유무를 판단하도록 제시하였는데, 인천 및 화성 지역의 CPTU 데이터의 50% 이상이 부분배수 상태를 나타내는 $B_q$ < 0.3에 분포하였다. 또한, 강제치환 시공과정 중 부분배수 현상으로 인해 원지반의 강도증가 현상이 발생되어 설계 예상 치환깊이보다 훨씬 작은 실측값이 얻어진다는 관점에서 실측 치환깊이와 동일한 값이 얻어지도록 원지반의 지지력에 대한 역해석을 수행하였다. 그 결과, 소성지수가 감소할수록 내부마찰각이 커지는 경향을 나타내며, 내부마찰각(${\varphi}^{\prime}$)이 $2{\sim}7^{\circ}$의 범위에서 분포하는 것으로 분석되었다.

목단피로부터 항균활성 성분의 분리 (Isolation of Antimicrobial Components from Moutan Cortex)

  • 권오근;김성환;천병열;박채규;손건호
    • 생약학회지
    • /
    • 제30권3호
    • /
    • pp.340-344
    • /
    • 1999
  • To evaluate antimicrobial activity of Moutan cortex the compounds isolated from $CHCl_3$ and EtOAc fractions of Moutan cortex were subjected to eight pathogenic strains. Benzoic acid, witch was identified from the $CHCl_3$ fraction, had MICs with $625{\sim}1,250\;{\mu}g/ml$ against all of the strains tested. Methyl gallate, p-hydroxy benzoic acid, gallic acid and $1,2,3,4,6-penta-O-gallyol-{\beta}-D-glucose$, which were identified from the EtOAc fraction, showed the antimicrobial activity, and the methyl gallate had the widest antimicrobial activity with MICs of $625{\sim}5,000\;{\mu}g/ml$ against all strains tested. p-Hydroxy benzoic acid showed MICs of $1,250{\sim}2,500\;{\mu}g/ml$ against all of the strains tested except C.albicans. Gallic acid had the best antimicrobial activities with MICs against the Shigella dysenteriae and Streptococcus mutans-strains of 78.1 and $312.5\;{\mu}g/ml$, respectively, but not against the C. albicans. And $1,2,3,4,6-penta-O-gallyol-{\beta}-D-glucose$ had the best antimicrobial activitie with MICs against the B. cereus, Staph. epidermidis and C. albicans strains of 39.1, 39.1 and $156.3\;{\mu}g/ml$, respectively, but not against the E. coli and Shig. Dysenteriae.

  • PDF