• 제목/요약/키워드: Sal I

검색결과 96건 처리시간 0.034초

Pseudomonas syringe pv. phaseolicola로 부터 제한효소의 분리정제 및 특성 (Purification and Characterzation of a Restriction Endonuclease from Pseudomonas syringae pv.phaselicola)

  • 배무;이은영
    • 한국미생물·생명공학회지
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    • 제22권5호
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    • pp.485-490
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    • 1994
  • A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

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디옥소몰리브덴 착물의 합성과 그 성질에 대한 연구 (Study on Formation and Properties of Dioxomolybdenum Complexes)

  • 오상오;구본권
    • 대한화학회지
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    • 제30권5호
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    • pp.441-448
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    • 1986
  • 몰리브덴산암모늄 수용액에서 적절한 살리실알데히드의 메탄올 혼합용액으로부터 디옥소비스(치환-살리실알데히다토) 몰리브덴늄(VI), $MoO_2(X-sal)_2(X=H,\;5-CH_3)$착물을 합성하고, 이들 각각의 화합물과 다양한 일차아민의 반응에 의하여 디옥소비스(치환-살리실알디미나토) 몰리브덴늄(VI)착물, $MoO_2(X-sal-N-R)_2,\;(R=C_6H_5,\;p-F-C_6H_4,\;m-Cl-C_6H_4,\;p-I-C_6H_4$$p-C_2H_5-C_6H_4)$을 합성하였다. 이들 착물은 모두 $900{\sim}940cm^{-1}$ 부근에서 ${\upsilon}_{Mo}=0$에 기인한 두 개의 강한 흡수띠가 관찰되었고, 핵자기공명스펙트럼에서 N=CH양성자에 대한 한 개의 시그날이 8.9ppm부근에서 나타났다. 이는 이들 착물이 $cis-MoO_2$기를 가진 6배위팔면체 착물임을 나타낸다. 질량분석의 결과로부터, $MoO_2$ : ligand의 결합비가 1:2임을 확인하였고, $MoO_2(5-CH_3-sal-N-R)_2$착물에 대한 분해과정을 조사하였다. 전자흡수스펙트럼에서 $N{\to}Mo$$O{\to}Mo$에 해당하는 전하이동전이는 $29,000cm^{-1}$$32,000cm^[-1}$부근에서 각각 일어났다. 한편, 디메틸포름아미드에서 측정한 이들 착물에 대한 몰전도도로부터 이들 착물이 비이온성 물질임을 확인하였다.

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Cloning and Expression in E. coli of the HOPDA Hydrolase Gene from Pseudomonas sp. P20

  • Lim, Jong Chul;Chae, Jong Chan;Kim Youngsoo;Kim, Hyong Bai;Kim Chi Kyung
    • Journal of Microbiology
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    • 제34권4호
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    • pp.349-354
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    • 1996
  • Pseudomonas sp. P20 is a natural isolate which is capable of degrading biphenyl and 4-chlorobiphenyl. From a clone of pCK1022 harboring pcbCD genes of Pseudomonas sp P20, a pcbD gene encoding 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoic acid (HOPDA) hydrolase was subcloned in Escherichia coli XL-1-Blue by using pBluescript SK(+) vector. The 2.8-kb HindII fragment harboring the pcbD gene cloned in pCK 1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 had a single site for each of XhoI, SalI, BstXI, and XbaI restriction enzymes. Escherichia coli CK1024 carrying pCK0124 degraded HOPDA to benzoate and 2-hydroxypenta-2, 4-dienoate by HOPDA hydrolase encoded by pcbD gene as effectively as E coli CK 1022 HARBORING pcbCD genes.

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Mitigating $CH_4$ Emissions in Semi-Aerobic Landfills: Impacts of Operating Conditions on Abundance and Community Structure of Methanotrophs in Cover Soils

  • Li, Huai;Chi, Zi-Fang;Lu, Wen-Jing;Wang, Hong-Tao
    • Journal of Microbiology and Biotechnology
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    • 제23권7호
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    • pp.993-1003
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    • 2013
  • Methanotrophs are the most important sink of $CH_4$, which is a more highly potent greenhouse gas than $CO_2$. Methanotrophic abundance and community diversity in cover soils from two typical semi-aerobic landfills (SALs) in China were detected using real-time polymerase chain reaction (real-time-PCR) and denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes, respectively. Real time-PCR showed that Type I methanotrophs ranged from $1.07{\times}10^6$ to $2.34{\times}10^7$ copies/g soil and that of Type II methanotrophs from $1.51{\times}10^7$ to $1.83{\times}10^8$ copies/g soil. The ratio of Type II to Type I methanotrophic copy numbers ranged from 5.61 to 21.89, indicating that Type II methanotrophs dominated in SAL. DGGE revealed that Type I methanotrophs responded more sensitively to the environment, changing as the community structure varied with different soil types and locations. Methylobacter, Methylosarcina, and Methylomicrobium for Type I, and Methylocystis for Type II were most prevalent in the SAL cover layer. Abundant interflow $O_2$ with high $CH_4$ concentration in SALs is the reason for the higher population density of methanotrophs and the higher enrichment of Type II methanotrophs compared with anaerobic landfills and other ecosystems, which proved a conclusion that increasing the oxygen supply in a landfill cover layer would greatly improve $CH_4$ mitigation.

Reevaluation of the Necessity of Iron Injection to Newborn Piglets

  • Yu, I.T.;Lin, J.;Wu, J.F.;Yen, H.T.;Lee, S.L.;Yang, T.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권1호
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    • pp.79-83
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    • 2002
  • The growth and immune responses to endotoxin lipopolysaccharide (LPS) challenge ( $20{\mu}g/kg$) of piglets with and without a iron dextran injection (Fe, 200 mg/head) two days after birth are compared. Sixty-four newborn piglets from eight litters were allocated randomly to one of four treatments. The control received no iron dextran and only saline (Sal) injection on the second and fifteenth day after birth (Sal-Sal). The remaining three groups received Fe-Sal, Sal-LPS, Fe-LPS treatments respectively. On fifteen days of age, blood samples of piglets were taken at 0 h, 1 h, 2 h and 4 d after saline or LPS injection to determine immune functions and blood characteristics. The trial terminated when the pig reached 56 days and the average daily gain of piglets was then measured. Daily gain, serum immunoglobulin G (IgG) concentration and red blood cell counts did not vary significantly among the four groups at any measuring times. Serum tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) concentration increased sharply 1 h after LPS injection. However, iron injection did not change TNF-${\alpha}$ concentration responds to LPS injection. White blood cell counts of two LPS injection groups were significantly lowered 1 h following the injection. In contrast, serum lactoferrin concentration had increased significantly 1 and 2 h postinjection. Furthermore, iron injection produced no further effects on these two criteria. Iron injection increased the hemoglobin (Hb) concentration of piglets at any measuring time, and LPS injection lowered Hb concentration. In conclusion, a 200 mg/head of iron dextran injection on the second day after birth increased Hb concentration, had no detrimental effect on the immune responses and growth of piglets. Moreover, if creep feed (175 mg Fe/kg feed) is provided from d 7 after birth, the Fe-injection does not contribute to overall performance of piglets and may not be a necessity in practice.

지방형 여성형유방증에 대한 새로운 분류와 1000증례 (The new classification for fatty-type gynecomastia (lipomastia) and 1000 cases review)

  • 윤상엽;강민구
    • Archives of Plastic Surgery
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    • 제36권6호
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    • pp.773-778
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    • 2009
  • Purpose: The authors propose the new classification of fatty - type gynecomastia(lipomastia) which can serve as a guide for modifying the periareolar technique. Methods: A retrospective analysis was made of 1000 cases of lipomastia operated on in the last 17 months. The extent of the clinical result, the technique employed, and the complications were observed. On the basis of this review the authors observed that at grade I(fat component < 50 ml, fibroglandular component < 3 g each breast), flattening of the thorax can be achieved by means of stab incision, ultrasound - assisted lipectomy(UAL), scavenging suction - assisted lipectomy(SAL) and tissue shaving. At grade II(50 < < 150 ml, 3 < < 5 g), stab incision, UAL, SAL and pull - out method(POM) using small curved scissors. At grade III(150 < < 300 ml, 5 < < 15 g and prominent inframammary fold(IMF)), minimal incision (5 - 6 mm), UAL, SAL and POM using small angulated scissors, and blunting IMF. At grade IV (300 < < 500 ml, 15 < < 30 g, and glandular ptosis), minimal incision (5 - 6 mm), UAL, SAL, fibroglandular excision using small angulated scissors, cutting IMF and fixation of nipple - areola complex(NAC) becomes necessary. At grade V (> 500 ml, > 30 g and ptosis), small incision (7 - 8 mm), UAL, SAL, fibroglandular excision using large angulated scissors, cutting IMF, upper repositioning of NAC and delayed circumareolar skin reduction or chest lifting becomes necessary. Results: The complications were minimal but there were hematoma (n = 7), infection (n = 3) and hypertrophic scar (n =13). Almost patients were satisfied with the outcome. Conclusion: This simple classification may help in choosing the most suitable treatment, thus avoiding insufficient or invasive treatments and undesirable scars.

Fusarium section Liseola 균주들에서 rDNA Intergenic Spacer 부위의 PCR-RFLP 분석 (PCR-RFLP Analysis of Ribosomal DNA Intergenic Spacer Region in Fusarium section Liseola.)

  • 이경은;최영길;민병례
    • 미생물학회지
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    • 제38권1호
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    • pp.7-12
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    • 2002
  • Fusarium section Liseola에 속하는 균주들의 rDNA IGS 부위를 중폭하고 여러개의 제한 효소로 처리하였다. 증폭된 IGS 부위의 길이는 F. moniliforme 12 만이 약 2.9 Kb 이고 나머지 균주들은 모두 약 2.6 Kb였다. 제한 효소 EcoRI, HincII, SalI, PstI 등은 ICS 부위를 절단하여 11균주들에서 9 haplotypes을 확인할 수 있었다. 본 연구에서의 Section Liseola에 속하는 균주들에 대한 결과에 앞서 연구되어진 Section Elegans에 속하는 균주들의 결과를 종합하여 dendrogram을 그렸을 때, IGS 부위를 종내에서와 마찬가지로 종간, section 간의 관계를 밝힐 수 있는 가능성을 제시하여 주었다.

알카리 내성 Bacillus속 Promoter의 Cloning (Cloning of Promoters from Alkali-tolerant Bacillus sp.)

  • 유주현;구본탁;공인수;정용준;박영서
    • 한국미생물·생명공학회지
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    • 제16권2호
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    • pp.126-130
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    • 1988
  • 토양에서 분리한 알카리 내성 Bacillus sp. YA-14 의 promoter를 Bacillus promoter probe vector인 pPL703을 이용하여 Bacillus subtilis내에 cloning 하였다. 얻어진 형질전환체 중 promoter 활성이 가장 높은 균주의 CAT 비활성은 8.07로 expression vector인 pPL708의 CAT 비활성보다 2.5배 이상 높았으며 대수 증식기가 끝난 이후에 그 활성이 급격하게 증가하였다. 재조합 plasmid내의 삽입 DNA 단편은 그 크기가 2.8kb이고 제한효소 BamHI, Sal I 인식부위가 각각 한군데 존재하였다.

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Spatial Patterns of Methane Oxidation and Methanotrophic Diversity in Landfill Cover Soils of Southern China

  • Chi, Zi-Fang;Lu, Wen-Jing;Wang, Hong-Tao
    • Journal of Microbiology and Biotechnology
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    • 제25권4호
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    • pp.423-430
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    • 2015
  • Aerobic CH4 oxidation is an important CH4 sink in landfills. To investigate the distribution and community diversity of methanotrophs and link with soil characteristics and operational parameters (e.g., concentrations of O2, CH4), cover soil samples were collected at different locations and depths from the Mengzi semi-aerobic landfill (SAL) in Yunnan Province of southern China. Specific PCR followed by denaturing gradient gel electrophoresis and realtime PCR were used to examine methanotrophs in the landfill cover soils. The results showed that different locations did harbor distinct methanotroph communities. Methanotrophs were more abundant in areas near the venting pipes because of the higher O2 concentrations. The depth of 20-25 cm, where the ratio of the CH4 to O2 was within the range from 1.3 to 8.6, was more conducive to the growth of CH4-oxidizing bacteria. Type II methanotrophs dominated in all samples compared with Type I methanotrophs, as evidenced by the high ratio of Type II to Type I methanotrophic copy numbers (from 1.76 to 11.60). The total copy numbers of methanotrophs detected were similar to other ecosystems, although the CH4 concentration was much higher in SAL cover soil. Methylobacter and Methylocystis were the most abundant Type I and Type II methanotrophs genera, respectively, in the Mengzi SAL. The results suggested that SALs could provide a special environment with both high concentrations of CH4 and O2 for methanotrophs, especially around the vertical venting pipes.

대장균 내에서의 Bdi I Methylase 유전자의 클로닝과 발현 (Cloning and Expression of the Bdi Methylase Gene in E. coli)

  • 전희숙;김용석;최경래;노현모
    • 미생물학회지
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    • 제25권1호
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    • pp.40-45
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    • 1987
  • B Brevibacterium divaricatum FERM 5948 균주로부터 Bdi I RIM 체계에 속하는 BdiI methylase 유천자를 클로닝하여 발현을 조사하였다. Bdi I methylase 유전자의 클로닝을 위해 pBR 322의 EcoRI, BamHl, Sal I 3 군데의 클로닝 site를 이 용했고 1 차 형질전환후 나온 플라스미드를 BdiI으로 자른 뒤 ligation 시키지 않고 형질전환시키는 방법을 이용하였다. 유전 자을 가지는 행질전환체의 선별은 Bdi I methylase에 의해 수정된 채조합 플라스미드는 BdiI 제한효소에 방호된다는 것에 기 초하여 선별하였는데 5.6kb의 EcoRI insert DNA를 가지는 pBDIM 116이 Bdil methylase 유전자플 가지는 것으로 판명 되었다. pBDIM 11&을 가지는 숙주셰포에서 추출한 추출용액에는 S-adenosylmethionine이 있으면 BdiI의 인지부위인 A ATCGAT에만 특정한 methylase 활성이 측정되였다. 11개의 제한효소를 이용하이 제한효소지도를 작성하였고, BdiI r restriction -modification 체계에 관해서 도 논의하였다.

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