• 제목/요약/키워드: agarose

검색결과 615건 처리시간 0.022초

Improvement of a Unified Saccharification and Fermentation System for Agaro-bioethanol Production in Yeast

  • Lee, So-Eun;Kim, Yeon-Hee
    • 한국미생물·생명공학회지
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    • 제48권1호
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    • pp.32-37
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    • 2020
  • We improved on a unified saccharification and fermentation (USF) system for the direct production of ethanol from agarose by increasing total agarase activity. The pGMFα-NGH plasmid harboring the NABH558 gene encoding neoagarobiose hydrolase and the AGAG1 and AGAH71 genes encoding β-agarase was constructed and used to transform Saccharomyces cerevisiae 2805. NABH558 gene transcription level was increased and total agarase activity was increased by 25 to 40% by placing the NABH558 gene expression cassette upstream of the other gene expression cassettes. In the 2805/pGMFα-NGH transformant, three secretory agarases were produced that efficiently degraded agarose to galactose, 3,6-anhydro-L-galactose (AHG), neoagarobiose, and neoagarohexaose. During the united cultivation process, a maximum of 2.36 g/l ethanol from 10 g/l agarose was produced over 120 h.

Evaluation of Several Parameters of in situ Polymerase Chain Reaction (ISPCR) to Reduce the Leakage of Amplificants from Cells

  • Lee, Jae-Yung;Auh, Chung-Kyoon;George W. Jordan
    • Journal of Microbiology
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    • 제40권1호
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    • pp.70-76
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    • 2002
  • Proviral DNAs from HIV-1-infected CD4+ T cells (Molt/LAV cells) were amplified and detected in infected individual cells using polymerase chain reaction and in rifu hybridization. In this in situ PCR, three parameters were considered to achieve effective amplification and retention of amplificants inside the cells by making high molecular weight PCR products intracellularly, forming agarose matrix against the cells, and maintaining the appropriate PCR temperature profile. Over the cycles of ampliHcationl tailed primers with complementary overhanging sequences at their 5' sides manufactured high molecular weight products by using short primary products as a repeating unit. Agarose matrix could prevent the diffusion of the amplificants from the cells. Use of Thermanox coverslip inside the PCR tube offered target cells a similar temperature profile to that of conventional PCR in solution.

Evaluation of Genetic Diversity among Korean Wild Codonopsis lanceolata by Using RAPD

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    • 한국자원식물학회지
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    • 제10권3호
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    • pp.258-264
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    • 1997
  • The introduction of molecular biology methodologies to plant improvement programs offers an invaluable opportunity for extensive germplasm characterization. We have applied the developed technique of random amplification of polymorphic DNA(RAPD)to the analysis of evaluating genetic diversity among Korean wild Codonopsis lanceolata. A total of 340 polymorpic hands were gernerated on agarose- and polyacrylamide-gel by 19 primers of abitrary sequence. grouped by cluster analysis using sample matching coefficients of similarity. Among of the samples. the minimum genetic distance value was obtained between sample no. 1(Girisan) and no. 2(Girisan), and the largest value between sample no. 11(Sulaksan) and no. 17(Sulaksan).In separate cluster dendrograms based on agareose - and polyacryamide-gel. some differences were observed; In the case of agarose gel,41 samples could be devided into 7 groups at below about 0.44 level of distance. However they were divided into 6 gourps at below about 0.40 level of distance in the case of polyacrylamide gel. These results showed that polymophic data in agrose were not grouped to wild plant selected from each mountainous district except for wild plants selected from Sulaksan and Chiaksan. We believe that polyacrylamide-RAPD is a superior method for detecting DNA polymorphism compared to agarose-RAPD method.

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플렉서블 플라즈마 패치에서 발생되는 활성종이 다당류 표면에 미치는 영향 (Effect of Active Species Generated from Flexible Plasma Patch on Polysaccharide Surface)

  • 이유리;이승훈;김도근
    • 한국표면공학회지
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    • 제51권2호
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    • pp.133-137
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    • 2018
  • Plasma devices such as jets, pencils, and torches have been developed as new tools that help penetration of target agents and applied to plasma medicine. However, these devices cannot be used in a large area. Therefore, we introduced a flexible plasma device, which can be treated of large area and designed as bendable plasma. In additional, in vitro model based on agarose gel was prepared that can be show effectiveness in the depth of penetration. Plasma treatment conditions such as power, time and distance can be optimized on the agarose gel wound model. The chemical structure of changed polysaccharides was predicted due to reactive excited atoms and molecules, UV photons, charged particles and reactive oxygen and nitrogen species (RONS).

DNA Fingerprinting by Amplified Fragment Length Polymorphism Markers in Rainbow Trout(Oncorhynchus mykiss)

  • Yoon, Jong-Man;Park, Sang-Hoon
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2001년도 춘계 수산관련학회 공동학술대회발표요지집
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    • pp.559-560
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    • 2001
  • The objective of the present study was to analyze genetic variation and characteristics in rainbow trout(Oncorhynchus mykiss) using amplified fragment length polymorphism(AFLP) method as molecular genetic technique, to evaluate the usefulness of AFLP as genetic markers, and to compared the efficiency of agarose and polyacrylamide sequencing gels. The amplified products were performed by agarose and sequencing gel electrophoresis to detect AFLP band patterns, respectively. Using 9 primer combinations, total of 141 AFLP bands were produced, 108 bands(82.4%) of which were polymorphic in agarose gels. In sequencing gels, total of 288 bands were generated, and 220 bands (76.4%) were polymorphic. The level of bandsharing(BS) ranged from 0.18 to 0.32 for the 9 primer combinations tested, with a mean of 0.24. Consequently, AFLP markers of these rainbow trout could be used as genetic information such as species identification, genetic relationship or analysis of genome structure, and selection aids for genetic improvement of economically importment traits in fish species.

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해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특성 (Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3)

  • 김다솜;김종희;지원재
    • 한국미생물·생명공학회지
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    • 제49권3호
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    • pp.329-336
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    • 2021
  • Cellulophga sp. J9-3은 셀룰로스 분해능력을 갖으며, Flavobacteriaceae 과에 속하는 그람-음성 호기성 해양 세균이다. 또한, J9-3 균주는 고체 및 액체 배지에서 한천을 가수 분해 할 수 있으며, 배지에 첨가한 아가로스(agarose)에 의해 아가레이즈(agarase)의 생산이 현저하게 유도되는 특성을 보였다. Cellulophga sp. J9-3의 세포 배양액으로부터, 황산암모늄 침전 및 3 단계의 컬럼 크로마토그래피를 연속적으로 수행하여, 한 개의 agarase 단백질, AgaJ93을 순수하게 정제하였다. 정제된 AgaJ93은 아가로스에 대한 분해 활성이 가장 강하였으며, starch에 대해서도 아가로스 대비 약 22% 정도의 분해 활성을 나타냈다. AgaJ93은 아가로스를 분해하여 사이즈가 큰 올리고당 중간체를 경유하여, 최종산물로 네오아가로테트라오스와 네오아가로헥사오스까지 분해함을 확인하였으며, 이는 AgaJ93이 endo-type β-아가레이즈 임을 의미한다. AgaJ93은 pH 7.0, 35℃에서 최대 활성을 나타냈고, Co2+ 이온에 의해 6배 이상의 활성증가를 보였다. AgaJ93의 N-terminal sequence 분석 결과, AgaJ93은 Cellulophaga sp. W5C의 내열성 endo-type β-아가레이즈 Aga2와 82%의 상동성을 보였으나, 두 효소의 생화학적 특성이 달랐다. 따라서, AgaJ93은 기존에 보고된 β-아가레이즈 들과는 다른 신규의 아가레이즈 일 것으로 예상된다.

메칠머큐리젤에 분리된 리보핵산의 니트로셀루로스막으로 이동 (Transfer of RNA from Methylmercury-agarose Gel to Nitrocellulose Membrane)

  • 안정선
    • Journal of Plant Biology
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    • 제30권2호
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    • pp.109-116
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    • 1987
  • Effects of staining, buffer washing and denaturing agents on the transferrability of RNA fractionated on a methylmercury hydroxide-agarose gel to a nitrocellulose membrane were studied. Ethidium bromide staining and ammonium acetate buffer washing inhibited RNA transfer, while 3% HCHO and 0.5 M NaOH treatments stimulated transfer which was negated in the ammonium acetate buffer. Accordingly, maintenance of primary structure of RNA was proved to be essential for transferring RNA from the methylmercury hydroxideagarose gel to the nitrocellulose membrane.

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Klebsiella pneumonia로 부터 제한효소 Kpn I의 분리 (Isolation of kpn I restriction endonuclease from klebsiella pneumonia)

  • 이상철;이대실;유명희
    • 미생물학회지
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    • 제25권1호
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    • pp.73-79
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    • 1987
  • A restriction endonuclease, Kpn I has been isolated from Klebsiella pneumonia. Cells were broken by sonication. After ultracentrifugation the supernatant containing Kpn I activity was further purified by Sepharose-6B gel filtration, DEAD-Cellulose, Heparin-Agarose, and Aminohexyl-Agarose column chromatography. Final enzyme preparation was essentially free of contamination exonuclease and phosphatase, as judged by ligation-recut test. Total activity of the enzyme recovered from 10 grams of cells was $4.7\times 10^5$ units.

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Indica 벼의 원형질체들로부터 식물체 재분화 (Plant Regeneration from Protoplasts of Indica Rice)

  • 이성호;손영길;이수인;김주현;조무제
    • 한국작물학회지
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    • 제42권5호
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    • pp.615-625
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    • 1997
  • Indica 벼의 원형질체들로 부터 효과적인 식물체 재분화 방법을 개발하였다. 이 방법은 배형성 진탕 세포 배양체로 부터 나출된 원형질체들을 feeder cell들이 agarose에 embedded된 배지 표면에 놓인 filter membrane 위에 배양하는 것이다. Feeder cell로서 Lolium multiflorum 세포배양체를 사용했을 때가 Oryza ridleyi를 사용했을 때보다 효과적이었고, 원형질체 평탄효율은 세포 배양체 age에 따라 달랐지만 최고로 0.68% 까지 증가되었다. Carbohydrate source로서 maltose를 사용하거나 maltose와 sucrose를 1:1로 조합했을 때가 sucrose 단독으로 사용했을 때보다 식물체 재분화율이 증가되었고, 고농도의 agarose를 이용하여 원형질체로부터 유도된 캘러스를 dehydration시켰을 때 또한 재분화율이 괄목하게 증가되었다. 식물체 재분화율은 control 캘러스로부터 3.1∼30.6% 였지만 dehydration처리한 캘러스로부터는 30.7∼70.7%까지 증가되었다. 원형질체로 부터 유도된 식물체들은 형태적으로 정상이며 개화했다.

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