• 미생물학회지
• /
• 제32권3호
• /
• pp.198-201
• /
• 1994

Lactobacillus casei에 감염하는 bacteriophage J1 게놈의 cohesive end site (cos)의 염기배열을 결정하였다. 또한 환형 cos와 선형 J1 DNA의 왼쪽 말단 염기배열을 비교한 결과 terminase가 절단하는 위치는 다음과 같았다. 5'- GGTCGGCC$\downarrow$ -3' 3'- $\uparrow$CCAGCCGG -5' J1 게놈의 cohesive end는 3' 말단이 돌출되어 있으며 8개의 뉴클레오티드로 이루어져 있고 G+C 함유율이 87.5%이었다. cos 부위는 선형 DNA의 왼쪽 5' 말단 뉴클레오티드의 위치를 +1로 정하였을 때 -33부터 +25까지 대칭이었다. 지금까지 보고된 phage들의 cos 부위 사이에 상동성은 발견되지 않았다.

• 한국수자원학회:학술대회논문집
• /
• 한국수자원학회 2011년도 학술발표회
• /
• pp.17-17
• /
• 2011

As a method of countermeasure to bed degradation and armoring phenomena of bed material in the downstream area of dam reservoirs, sediment augmentation (replenished sediment) has been carried out in many Japanese rivers. In general, bed of the replenished sediment site is composed of rocks, because the site is located in the downstream area of the dams and sediment supply is very small. Bed deformation process has been researched by many researchers. As a method of countermeasure to bed degradation and armoring phenomena of bed material in the downstream area of dam reservoirs, sediment augmentation (replenished sediment) has been carried out in many Japanese rivers. In general, bed of the replenished sediment site is composed of rocks, because the site is located in the downstream area of the dams and sediment supply is very small. Bed deformation process has been researched by many researchers. However, most of them can treat movable bed only and cannot be applied to the bed deformation process of sediment on rocks. If the friction angle between the sediment and the bed surface is assumed to be the same as the friction angle between the sediment and the sediment, sediment transport rate must be smaller without sediment deposition layer on the rocks. As a result, the reproduced bed geometry is affected very well. In this study, non-equilibrium transport process of non-cohesive sediment on rigid bed is introduced into the horizontal two dimensional bed deformation model and the model is applied to the erosion process of replenished sediment on rock in the Nakagawa, Japan. Here, the Japanese largest scale sediment augmentation has been performed in the Nakagawa. The results show that the amounts of the eroded sediment and the remained sediment reproduced by the developed numerical model are $56300m^3$ and $26800m^3$, respectively. On the other hand, the amounts of the eroded sediment and the remained sediment measured in the field after the floods are $56600m^3$ and $26500m^3$, respectively. The difference between the model and field data is very small. Furthermore, the bed geometry of the replenished sediment after the floods reproduced by the developed model has a good agreement with the measured bed geometry after the floods. These results indicate that the developed model is able to simulate the erosion process of replenished sediment on rocks very well. Furthermore, the erosion speed of the replenished sediment during the decreasing process of the water discharge is faster than that during the increasing process of the water discharge. The replenished sediment is eroded well, when the top of the replenished sediment is covered by the water. In general, water surface level is kept to be high during the decreasing process of the discharge during floods, because water surface level at the downstream end is high. Hence, it is considered that the high water surface level during the decreasing process of the water discharge affects on the fast erosion of the replenished sediment.

• 미생물학회지
• /
• 제32권1호
• /
• pp.91-95
• /
• 1994

토양에서 분리된 Enterobacter agglomerans CBNU45는 type II 제한효소인 EagBI을 생산하고 있음을 발견하였다. EagBI을 DEAE-cellulose, phosphocellulose P11, hydroxylapatite column chromatography를 거쳐 부분 정제하여 그 특성을 알아보았다. EagBI은 여섯개의 염기배열 5’-CGAT${\downarrow}$CG-3’을 인식하고 T 와 C 사이를 절단하여 두개의 염기가 3’-말단쪽으로 돌출된 cohesive end를 형성하였다. EagBI의 반응 최적조건은 10mM Tris-HCl(pH 7.8), 6~10mM $MgCl_2$, 37${\circ}C$이었으며 NaCl이 없는 반응 완충용액에서 가장 좋은 활성을 보였다. EagBI은 $dam^-$와 $dam^+$ 메칠화 DNA도 절단할 수 있으며 65${\circ}C$에서 10분 동안 열처리하였을 때 효소의 활성을 상실하였다. 따라서 EagBI은 PvuI의 isoschizomer이나 NaCl 요구성과 열안정성에서 PvuI보다 편리한 제한효소로 확인되었다.

• 대한바이러스학회지
• /
• 제26권1호
• /
• pp.77-90
• /
• 1996

Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.