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RISS 인기검색어
- 검색키워드
- 저자 : 고용송 ( Yong Song Gho ) (검색결과 5 건)
- 무료
- 기관 내 무료
- 유료
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Purification and Characterization of 3-Methyladenine DNA Glycosylase from Bacillus Subtilis
고용송,한범희,양철학,Gho, Yong-Song,Han, Bom-Ie,Yang, Chul-Hak 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.3
알킬화된 DNA로부터 주로 3-메칠아데닌을 제거하는 유발성 효소를 고초균으로부터 DEAE-셀루로오즈와 DNA-셀루로오즈 크로마토그래피법을 사용하여 부분적으로 분리하고 이를 3-메칠아데닌 DNA 글라이코실라아제로 명명하였다. 이 효소는 N-메칠-N-니트로소유리아로 메칠화된 DNA 기질로부터 3-메칠아데닌과 3-메칠구아닌을 제거하였다. 이 때 3-메칠아데닌은 7-메칠구아닌 보다 30배나 빨리 제거되었다. 이 효소는 $O^6$-메칠구아닌은 제거시키지 못하였다. 이 효소의 활동도는 3-메칠아데닌이나 7-메칠구아닌에 의해 방해되지는 않았으며 스퍼미딘에 의해 그 활동도가 증가되었다. 이 효소의 분자량은 SDS 아크릴아미드 젤 전기영동법에 의하면 30,000 정도이었고, 셰파덱스 G-100의 젤 여과법에 따르면 23,000으로 나타났다. 효소의 등전점은 등전초점화 콜럼에서 4.7로 나타났다. 두개의 3-메칠아데닌 DNA 글라이코실라아제가 존재하는 대장균과는 달리 고초균에는 한 형태의 효소만 존재함을 확인하였다. 또한 고초균 3-메칠아데닌 DNA 보다 겹가닥 알킬화된 DNA를 선호하였다. 기질을 알킬화된 DNA를 사용했을 때 $K_m$값은 $5.15{\times}10^{-8}M$이었다. 1 mM의 EDTA 존재하에 효소의 활동도는 약간 감소하였고 2mM $Mg^{2+}$와 $Ca^{2+}$의 존재하에 베이스의 제거가 증가되는 한편 2mM의 $Co^{2+}$와 $Mn^{2+}$은 효소의 활동도를 방해하였다. 이 효소반응의 pH 적정점은 pH6.7-7.5이었고 온도에 대해 특히 민감하였고 $37^{\circ}C$에서 예민한 적정점을 나타내었다. 이 효소는 썰퍼하이드릴 그룹에 반응하는 N-메칠말레이미드와 p-클로로머큐리벤조에 의해 불활성화되었다. An inducible enzyme which releases primarily 3-methyladenine from alkylated DNA was partially purified from Bacillus subtilis by using DEAE-cellulose and DNA-cellulose chromatography and designated as 3-methyladenine DNA glycosylase. It released 3-methyl-adenine and 7-methylguanine from DNA methylated with N-methyl-N-nitrosourea (MNU). 3-methyladenine (3-meA) was released 30 times faster than 7-methylguanine (7-meG). No detectable amount of $O^6$-methylguanine ($O^6$-meG) was released. The glycosylase activity was inhibited by neither free 3-meA nor 7-meG but was stimulated by spermidine. Apparent molecular weight was 30,000 on SDS polyacrylamide gel electrophoresis and 23,000 on Sephadex G-100 gel filtration. Its pI was 4.7 on column electrofocusing. Unlike the 3-methyladenine DNA glycosylases in Escherichia coli, no evidence for the presence of two different enzymes was found. The enzyme exhibited a preference for double-stranded alkylated DNA to the single stranded one. Apparent $K_m$ for substrate DNA was $5.15{\times}10^{-8}M$. Addition of 1 mM EDTA slightly decreased the activity. Addition of 2 mM $Mg^{2+}$ and $Ca^{2+}$ stimulated base release, while 2 mM $Co^{2+}$ and $Mn^{2+}$ inhibited it. Optimum pH for the enzyme reaction was pH 6.7-7.5. The enzyme activity was quite sensitive to temperature and had a sharp optimum at $37^{\circ}C$. The enzyme was inactivated by N-ethylmaleimide and p-chloromercuribenzoate which was known as a sulfhydryl reagents.
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고초균에서 3 - 메칠아데닌 DNA 글아이코실라아제의 분리 및 특성
고용송,한범희,양철학 ( Yong Song Gho,Bom He Han,Chul Hak Yang ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.3
An inducible enzyme which releases primarily 3-methyladenine from alkylated DNA was partially purified from Bacillus subtilis by using DEAE-cellulose and DNA-cellulose chromatography and designated as 3-methyladenine DNA glycosylase. It released 3-methyl-adenine and 7-methylguanine from DNA methylated with N-methyl-N-nitrosourea (MNU). 3-methyladenine (3-meA) was released 30 times faster than 7-methylguanine (7-meG). No detectable amount of O^6-methylguanine (O^6-meG) was released. The glycosylase activity was inhibited by neither free 3-meA nor 7-meG but was stimulated by spermidine. Apparent molecular weight was 30,000 on SDS polyacrylamide gel electrophoresis and 23,000 on Sephadex G-100 gel filtration. Its pI was 4.7 on column electrofocusing. Unlike the 3-methyladenine DNA glycosylases in Escherichia coli, no evidence for the presence of two different enzymes was found. The enzyme exhibited a preference for double-stranded alkylated DNA to the single stranded one. Apparent K_m for substrate DNA was 5.15×10^(-8)M. Addition of 1 mM EDTA slightly decreased the activity. Addition of 2 mM Mg^(2+) and Ca^(2+) stimulated base release, while 2 mM Co^(2+) and Mn^(2+) inhibited it. Optimum pH for the enzyme reaction was pH 6.7-7.5. The enzyme activity was quite sensitive to temperature and had a sharp optimum at 37℃. The enzyme was inactivated by N-ethylmaleimide and p-chloromercuribenzoate which was known as a sulfhydryl reagents.
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여창섭(Chang Sub Yeo),한규연(Kyu Yeon Han),고용송(Yong Song Gho),이상준(Sang Joon Lee) 대한기계학회 2005 대한기계학회 춘추학술대회 Vol.2005 No.11
To analyze in-vivo blood flow characteristics in a chicken embryo, in-vivo experiment was carried out using micro- PIV technique. Because endothelial cells in blood vessels are subject to shear stressof blood flow, it is important to get velocity field information of the placental blood flow. The flow images of RBCs were obtained with a spatial resolution of 20 20㎛ in the whole blood vessels. The blood in a branched vessel merged smoothly without any flow separation into the main blood vessel with the presence of a slight bump. This in-vivo micro-PIV measurement technique can be used as a powerful tool in various blood flow researches.
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