• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.204-204
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• 2022

Transformant construction using protoplasts requires less sample preparation time than particle bombardment and Agrobacterium-mediated transfection. There are two protoplast transfection methods: the PEG-mediated transfection method and the Lipofectamine transfection method. When Lipofectamine is mixed with DNA, Lipofectamine surrounds DNA like a cell membrane because of the positive charge of Lipofectamine. The Lipofectamine-DNA complex makes DNA insertion into cells easier. Fectin has similar functions to lipofectamine and is less expensive than lipofectamine. The 3D-fectin technology has been highlighted in animal cell transfection. Therefore, we performed PEG-mediated transfection, Lipofectamine transfection, and 3D-pectin transfection with a GFP construct. Protoplasts were isolated using the first leaf of "Bobwhite" after 4 hours of incubation in an isolation Buffer (cellulase + macerozyme). Protoplasts transformed by each method were cultured for 48 hours, and then GFP fluorescence expression was confirmed under confocal microscopy. GFP signals were detected in PEG-mediated transfection and Lipofectamine transfection. And the GFP signals were also detected in protoplasts to which 3D-fectin technology was applied, suggesting that 3D-fectin technology can be used for plant protoplast transfection.

• 한국버섯학회지
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• 제20권3호
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• pp.178-182
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• 2022

버섯의 오랜 역사에도 불구하고 버섯의 유전적 기능과 분자유전학을 응용한 신품종 개발에 대한 연구는 크게 부족한 상황이다. 그러나 최근 유전자 가위인 CRISPR/Cas를 이용한 새로운 유전자 교정 기술이 개발됨에 따라 버섯 연구에서 이 기술을 이용한 다양한 시도가 이루어지고 있다. 특히 선택의 용이성을 위해 외래 유전자 삽입 없이도 고효율로 유전자 편집이 가능한 RNPs를 활용한 연구가 활발히 진행되고 있다. 그러나 RNPs는 원형질체의 세포막을 통과하기에 Cas9이 너무 거대하고 guide RNA가 쉽게 파괴된다는 단점을 가지고 있다. 이러한 단점을 극복하기 위하여 세포막 통과에 용이한 미네랄 성분인 CaP와 PAA를 조합하여 Nanoparticle을 형성함으로써 극복하고자 했다. 표고버섯 단핵 균주인 산조705-13을 이용하여 원형질체 분리에 적합한 Osmotic buffer를 찾기 위하여 0.6M과 1.2M의 Sucrose, Sorbitol, Mannitol, KCl을 처리하였고 그 결과 0.6M Sucrose가 가장 적합한 osmotic buffer임을 확인하였다. 또한 CaP으로 RNPs와 Nanoparticle 복합체를 형성하고 이 복합체가 RNase A로부터 RNPs의 기능을 온전히 보호하는 것을 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제4권2호
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• pp.102-107
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• 1994

The $\beta$-1,4-endoglucanase gene from Actinomyces sp. 40 was cloned into Escherichia coli DH5$\alpha$ with pUC19. Chromosomal DNA from Actinomyces sp. 40 was cleaved with the restriction enzyme Sau3AI and ligated into pUC19 for the transformation of Escherichia coli DH5$\alpha$. Positive clones of $\beta$-1,4-endoglucanase gene were detected as the clear zones on a medium supplemented with carboxymethylcellulose (CMC). This transformant possessed a single plasmid, designated pDS1, which contained the vector DNA and a 3.5 kilobase (kb) Sau3AI insertion fragment encoding endoglucanase. The size of the cloned fragment was reduced to 2.0 kb. The endoglucanase activity produced by the E. coli DH5$\alpha$ (pDS6) was higher than that of Actinomyces sp. 40 strain. The optimum pH and temperature of the cloned enzyme were pH 4.0$\sim$5.0 and 55$^{\circ}C$, respectively. The cloned enzyme was stable at 55$^{\circ}C$ or below and in buffer ranging from pH 4.0 to 7.0. The enzyme degraded CMC but did not degrade xylan, cellobiose, and methyl-umbelliferylcellobiopyranoside (MUC).

• 한국재료학회지
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• 제19권10호
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• pp.517-521
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• 2009

Si-C composite with hollow spherical structure was synthesized using ultrasonic treatment of organosilica powder formed by hydrolysis of phenyltrimethoxysilane. The prepared powder was pyrolyzed at various temperatures ranging from 900 to 1300 $^{\circ}C$ under nitrogen atmosphere to obtain optimum conditions for Li-ion battery anode materials with high capacity and cyclability. The XRD and elemental analysis results show that the pyrolyzed Si/C composite at 1100 $^{\circ}C$ has low oxygen and nitrogen levels, which is desirable for increasing the electrochemical capacity and reducing the irreversible capacity of the first discharge. The solid Si-C composite electrode shows a first charge capacity of $\sim$500 mAhg$^{-1}$ and a capacity fade within 30 cycles of 0.93% per cycle. On the other hand, the electrochemical performance of the hollow Si-C composite electrode exhibits a reversible charge capacity of $\sim$540 mAhg$^{-1}$ with an excellent capacity retention of capacity loss 0.43% per cycle up to 30 cycles. The improved electrochemical properties are attributed to facile diffusion of Li ions into the hollow shell with nanoscale thickness. In addition, the empty core space provides a buffer zone to relieve the mechanical stresses incurred during Li insertion.

• 정보처리학회논문지B
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• 제12B권6호
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• pp.661-670
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• 2005

인터넷 상에서의 멀티미디어 스트림을 제공하는 서비스가 활성화됨에 따라 서비스의 사용자도 크게 증가하였다. 이것은 네트워크 트래픽의 증가로 이어졌고, 결과적으로 스트림 재생시 불연속적인 재생과 영상 및 음원의 비동기화와 같은 문제를 발생시켰다. 이러한 문제를 해결하기 위해 안정적인 미디어 스트림의 재생을 보장하고 부가적으로 서비스 사용자와 미디어 간 상호대화를 할 수 있는 미디어 전달 방법이 필요하다. 기존의 관련 연구에서는 여러 가지 방법을 통하여 미디어간 동기화를 달성하고 있으나 상호대화라는 측면에서는 만족할 만한 해결책을 제시하지 못하고 있으며, 불연속적인 스트림의 재생에 대한 처리에도 문제점을 가지고 있다. 본 논문에서는 상호대화형 객체를 각 미디어 파일에 삽입하고, 이들 객체들이 서로의 정보를 이용할 수 있는 함수를 설계하여 동기화와 상호대화성 문제를 해결하며, 네트워크에 대한 의존성 때문에 발생하는 불연속적인 재생은 가변 버퍼를 이용함으로써 해결하였다.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.222-228
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• 2001

Vibrio metschnikovii strain RH530 is a non-pathogenic, industrially-important alkaline protease producer which has been isolated from wastewater. In this paper, we report on the transformation of this strain by using the method of electroporation. A field strength of $7.5\;kVcm^{-1}$ and $25\;{\mu}F$, and using a 0.2-cm cuvette, appeared to be the optimal conditions for electroporation of the cells with the recombinant pSBCm plasmid carrying the vapK alkaline protease gene and the ColE1 replicon. Cells were subjected to osmotic shock in order to remove extracelluar DNase, and adding 200 mM of sucrose to electroporation buffer cells showed an increased transformation efficiency. Maximum efficiency of transformation was obtained at an early exponential growth phase. Using all of the conditions mentioned above, we routinely obtained a transformation efficiency of more than $10^4{({\mu}g\;plasmid\;DNA)}^{-1}$. The stability of the plasmid pSBCm in V. metschnikovii RH530 was 25% after 18h of growth (27 generations) in the medium without antibiotic selection. The insertion of the par locus to the pSBCm increased the stability of the plasmid up to 42% without selective pressure. The increase in plasmid stability was accompanied by the increase in the productivity of alkaline protease in the recombinant V. metschnikovii strain RH530. Determining optimal conditions for the transformation of the industrially-important, nonpathogenic Vibrio strain, and the improvement of plasmid stability by introducing the par locus into the high copy number plasmid vector, will allow the development of procedures involved in the genetic manipulation of this strain, particularly for its use in the production of industrial enzymes such as alkaline protease.

• BMB Reports
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• 제34권4호
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• pp.293-298
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• 2001

Tryptic activation of the 130-kDa Bacillus thuringiensis Cry4B $\delta$-endotoxin produced protease-resistant products of ca. 47 kDa and ca. 21 kDa. The 21-kDa fragment was identified as the N-terminal five-helix bundle (${\alpha}1-{\alpha}5$,) which is a potential candidate for membrane insertion and pore formation. In this study, we constructed the recombinant clone over-expressing this putative pore-forming (PPF) fragment as inclusion bodies in Escherichia coli. The partially purified inclusions were composed of a 23-kDa protein, which cross-reacted with Cry4B antibodies, and whose N-terminus was identical to that of the 130-kDa protein. Dissimilar to protoxin inclusions, the PPF inclusions were only soluble when the carbonate buffer, pH 9.0, was supplemented with 6 M urea. After renaturation via a stepwise dialysis, the refolded PPF protein appeared to exist as an oligomer and was structurally stable upon trypsin treatment. Unlike the 130kDa protoxin, the refolded protein was able to release entrapped glucose from liposomes, and showed comparable activity to the full-length activated toxin, although it lacks larvicidal activity These results, therefore, support the notion that the PPF fragment that consists of ${\alpha}1-{\alpha}5$ of the activated Cry4B toxin is involved in membrane pore-formation.

• BMB Reports
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• 제34권5호
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• pp.402-407
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• 2001

Based on the currently proposed toxicity model for the different Bacillus thuringiensis Cry $\delta$-endotoxins, their pore-forming activity involves the insertion of the ${\alpha}4-{\alpha}5$ helical hairpin into the membrane of the target midgut epithelial cell. In this study, a number of polar or charged residues in helix 4 within domain I of the 65-kDa dipteranactive Cry11A toxin, Lys-123, Tyr-125, Asn-128, Ser-130, Gln-135, Arg-136, Gln-139 and Glu-141, were initially substituted with alanine by using PCR-based directed mutagenesis. All mutant toxins were expressed as cytoplasmic inclusions in Escherichia coli upon induction with IPTG. Similar to the wild-type protoxin inclusion, the solubility of each mutant inclusion in the carbonate buffer, pH 9.0, was relatively low When E. coli cells, expressing each of the mutant proteins, were tested for toxicity against Aedes aegypti mosquito-larvae, toxicity was completely abolished for the alanine substitution of arginine at position 136. However, mutations at the other positions still retained a high level of larvicidal activity Interestingly, further analysis of this critical arginine residue by specific mutagenesis showed that conversions of arginine-136 to aspartate, glutamine, or even to the most conserved residue lysine, also abolished the wild-type activity The results of this study revealed an important determinant in toxin function for the positively charged side chain of arginine-136 in helix 4 of the Cry11A toxin.

• 정보처리학회논문지D
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• 제8D권5호
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• pp.593-604
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• 2001

데이타베이스 공유 시스템(Database Sharing System:Dss)은 고성능의 트랜잭션 처리를 위해 제안된 구조이다. DSS에서 고속의 통신망으로 연결된 노드들은 별도의 메모리와 운영체제를 가지며, 데이타베이스를 저장하고 있는 디스크 모든 노드에 의해 공유된다. 빈번한 디스크 액세스를 피하기 위해 각 노드는 최근에 액세스한 데이타 페이지와 인덱스 페이지들을 자신의 메로리 버퍼에 캐싱한다. 일반적으로 B-트리 인덱스페이지들은 데이타 페이지에 비해 빈번하게 캐싱되고, Fetch, Fetch Next, 삽입, 그리고 삭제와 같은 복잡한 연산을 수행하므로, 높은 동시성을 지원하는 효율적인 캐쉬 일관성 기법이 필요하다. 본 논문에서는 DSS에서 B-트리 인덱스 페이지의 식별자와 리프 페이지의 PageLSN을 사용한 캐쉬 일관성 기법을 제안한다.

• 한국정보통신학회논문지
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• 제26권10호
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• pp.1537-1544
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• 2022

application specific integrated circuit (ASIC) 및 system on chip (SoC) 설계 시 디지털 회로는 클럭에 동기화되어 작동한다. 칩 설계 시, place & route (P&R)에서 설계 조건과 타이밍 조건, 클럭의 동기화 여부 등을 고려한다. P&R에서 클럭 경로에 대한 delay를 줄이기 위해, clock tree synthesis (CTS) 기법을 이용한다. 본 논문에서는 사전 클럭트리 합성 가능 여부 판단을 위한 shallow-CTS 알고리즘을 소개한다. 오픈 소스 Parser-Verilog를 사용하여 register transfer level (RTL) 합성가능한 Verilog를 파싱하여, Pre-CTS와 Post-CTS 단계를 진행하고, 가장 긴 clock path와 버퍼 삽입 전후의 표준편차를 비교하여 CTS의 정확도에 대해 분석한다. 본 논문에서 시간 투입이 많이 되는 licensed EDA tool을 사용하여 CTS 결과를 확인하지 않고, RTL 수준에서 사전 클럭 트리 합성 검증 방법을 제공하여 비용 및 시간문제를 감소할 수 있을 것으로 기대된다.