• 미생물학회지
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• 제26권3호
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• pp.162-166
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• 1988

Bacteriophage M13 DNAs carrying the wild type or base substituted SV40 DNA replication origins were used for replication assay. In vivo and in vitro assay with African green monkey cell line COS-1 showed that the replication of M13-SV40 recombinant DNAs was restricted like a pBR322 SV40 recombinant DNA(Lusky and Botchan, 1981). Furthermore, recombinant phage DNAs isolated from the transfected siminan cells subsequently show a reduced ability to retransform E. coli. But pATSV-W(Kim et al., 1988) was replicated in COS-1 cells normally. We think that a poison sequence may exist on bacteriophage M13 DNA like pBR322.

• Journal of Plant Biology
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• 제36권1호
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• pp.19-27
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• 1993

Effects of polyamines on DNA synthesis were studied in synchronized culture of Nicotiana tabacum L. When DFMO and DFMA, inhibitors of ornithine decarboxylase and arginine decarboxylase, respectively were initially applied to the cells, the polyamine contents were rapidly dropped and [methyl-3H] thymidine incorporation into DNA was markedly reduced during the early stage of culture period. Inhibition of DNA synthesis, however, was partially reversed when these inhibitors were applied simultaneously with putrescine. In addition, exogenous administration of putrescine also increased the DNA synthesis during the all over the culture period. In vitro activity of DNA polymerase from Nicotiana tabacum L. was promoted by increasing concentrations of polyamines in the reaction mixture. Maximal activity was shown at 5 mM putrscine, 0.5 mM spermidine and spermine, respectively. Lack of Mg2+ ion in the reaction buffer resulted in an inhibition of the enzyme activity by about 30%. The inhibition could not be completely reversed by application of polyamines at optimal concentrations. These results suggest that polyamines promote the DNA synthesis in vivo and in vitro by stabilizing the DNA-helix upon binding to negatively charged groups on DNA or increasing the activity of DNA polymerase in Nicotiana tabacum L.

• Journal of information and communication convergence engineering
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• 제10권2호
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• pp.210-214
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• 2012

A microtiter well plate DNA hybridization method using Mycobacterium kansasii-specific rpoB DNA probe (kanp) were evaluated for the detection of M. kansasii from culture isolates. Among the 201 isolates tested by this method, 27 strains show positive results for M. kansasii, but the other 174 isolates were negative results for M. kansasii. This result was consistent with partial rpoB sequence analysis of M. kansasii and the result of biochemical tests. The negative strains by this DNA-DNA hybridization method were identified as Mycobacterium tuberculosis (159 strains), Mycobacterium avim (5 strains), Mycobacterium intracellulare (8 strains), and Mycobacterium flavescens (2 strain) by rpoB DNA sequence analysis. Due to high sensitivity and specificity of this test result, we suggest that DNA-DNA hybridization method using rpoB DNA probes of M. kansasii could be used for the rapid and convenient detection of M. kansasii.

• Parasites, Hosts and Diseases
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• 제35권3호
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• pp.203-210
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• 1997

이질아메바 병원성 분리주에서 특이적으로 발현되는 mRNA를 동정하고자 differential display reverse transcription-polymerase chain reaction(DDRT-PCR)을 수행하여 병원성 특이 증폭산물을 확인하였다. 한국인에서 검출한 이질아메바 병원성 분리주 YS-27과 Entamoeba dispar분리주인 S 16으로부터 정제한 mRAN를 주형으로 11개의 arbitrary primer와 3개의 one base anchored $oligo-dT_{11}M$(M: A, C 또는 G)의 조합을 이용, DDRT-PCR을 실시한 결과 31개의 분획이 YS-27주에서만 증폭된 것으로 확인되었다. 이 331개 DNA 중 21개는 cysteine proteinase 유전자와 상동성을 나타내었다. YS-27주로부터 제작된 cDNA library를 나머지 DNA를 탐침으로 사용, 검색하여 최종 4개의 clone을 얻었다. 이 4개의 clone을 이용, immunoscreening을 수행한 결과, 이 clone들은 이질아메바 감염자 혈청과 양성반응을 나타내고 있었다.

• The Korean Journal of Physiology and Pharmacology
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• 제6권2호
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• pp.109-112
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• 2002

The signal pathways involved in the regulation of AP-1 DNA binding activity in long-term nicotine stimulated bovine adrenal medullary chromaffin (BAMC) cells have not been well characterized. To understand the involvement of second messengers in the regulation of AP-1 DNA binding activity, the present study was designed to define the time-course for inhibition of nicotine-induced responses by cholinergic antagonists, $Ca^{2+}$ and calmodulin (CaM) antagonists, and calcium/calmodulin-dependent protein kinase (CaMK) II inhibitor using electrophoretic mobility shift assay. Nicotine $(10{\mu}M)$ stimulation increased AP-1 DNA binding activity at 24 hr after treatment. Posttreatment with hexamethonium (1 mM) plus atropine $(1{\mu}M)$ (HA), nimodipine $(1{\mu}M),$ or calmidazolium $(1{\mu}M)$ at 0.5, 3, and 6 hr after the nicotine treatment significantly inhibited the AP-1 DNA binding activity increased by long-term nicotine stimulation. However, posttreatment with HA, nimodipine, or calmidazolium at 9 or 12 hr after the nicotine treatment did not affect the nicotine-induced increase of AP-1 DNA binding activity. The pretreatment of BAMC cells with various concentrations of KN-62 inhibited the increase of AP-1 DNA binding activity induced by nicotine in a concentration-dependent manner. KN-62 $(10{\mu}M)$ posttreatment beginning at 0.5, 3, or 6 hr after the nicotine treatment significantly inhibited the increase of AP-1 DNA binding activity. However, KN-62 posttreatment beginning at 9 or 12 hr after the nicotine treatment did not affect the increase of AP-1 DNA binding activity. This study suggested that stimulation (for at least 6 hr) of nicotinic receptors on BAMC cells was necessary for increase of AP-1 DNA binding activity, and activation of $Ca^{2+},$ CaM, and CaMK II up to 6 hr at least seemed to be required for the increase of nicotine-induced AP-1 DNA binding activity.

• 한국식품영양과학회지
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• 제12권4호
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• pp.305-309
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• 1983

수종(數種)의 핵산구성성분(核酸構成成分), 아미노산(酸), 요소유도체(尿素誘導體)와 같은 생리활성물질(生理活性物質) 및 함질소화합물(含窒素化合物)의 DNA에 대한 작용(作用)과 그 작용(作用)에 무기(無機)ion의 영향을 검토(檢討)하였다. PTU, Cys-SH는 금속(金屬) ion없이도 DNA 절단능력(切斷能力)을 가지고 있었고, Tyr, Phe, Trp는 $CuSO_4$존재(存在) 하(下)에서 약한 DNA damage 유기작용(誘起作用)이 있었으며 5mM 이상의 Cys-SH와 $500{\mu}M$ $CuSO_4$의 혼합액(混合液)은 강(强)한 DNA 절단작용(切斷作用)을 나타냈다. $Sn^{2+}$를 제외한 $500{\mu}M$의 무기염(無機鹽)은 DNA에 대한 절단능력이 없었으나, $Cu^{2+}$, $Ni^{2+}$, $Mn^{2+}$, $Zn^{2+}$, $Sn^{2+}$ 등은 수종(數種)의 아미노산(酸)의 DNA breaking action에 영향을 미치고 있음을 확인(確認)되었다.

• 한국수산과학회지
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• 제20권2호
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• pp.136-145
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• 1987

말똥 성게를 인공 수정시킨 후 column chromatography법으로 DNA polymerase $\alpha$를 분리 정제하였다. Sephadex G-200과 SDS polyacryamise gel electophoresis에 의한 DNA Polymerase $\alpha$의 분자량은 약 $137,000\~138,000$이였다. 효소활성의 최적 pH는 7.4였고, 칼슘이온 20mM, 나트륨이온 25mM에서 활성이 높았고, 마그네슘 이온은 10 mM 일 때 활성이 높았다. DNA polymerase $\alpha$는 N-ethylmaleimide, aphidicolin, cytosin $\beta-D-arabinofuranoside$ 5'-triphosphate (ara CTP)와 phosphonoacetic acid체 의하여 활성이크게 저하되었다.

• Journal of Microbiology and Biotechnology
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• 제28권9호
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• pp.1467-1472
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• 2018

In this study, DNA-templated copper nanoclusters (DNA-CuNCs) were used to detect exonuclease III (Exo III) activity, which is important for the diagnosis and therapy of several diseases. The results of this study showed that Exo III was affected by the concentrations of magnesium ions and sodium ions, and its optimal conditions for cleavage were $5mM\;Mg^{2+}$ and less than $25mM\;Na^+$. With a blunt-end DNA, more than 98% of DNA was digested by Exo III. As expected, with two or four cytosines in the terminal position of a 4-base overhanging DNA such as 5'-GGCC-3' and 5'-CCCC-3', there was little cleavage by Exo III compared with a blunt-end DNA.

• BMB Reports
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• 제32권5호
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• pp.492-496
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• 1999

We have investigated the biochemical properties of the herpes simplex virus type 1 (HSV-1) DNA polymerase without the UL42 protein (Pol), purified from insect cells infected with a recombinant baculovirus containing the UL30 gene. BSA and DTT have inhibitory effects on dAMP incorporation. Pol showed a greater turnover rate of steady-state single nucleotide incorporation at 12 mM $MgCl_2$ than at 2 mM $MgCl_2$. However, it showed a greater processivity of DNA synthesis at lower $MgCl_2$ concentration (1 mM, 2 mM) than at a higher $MgCl_2$ concentration (12.5 mM). These results are consistent with a slow DNA dissociation at lower $MgCl_2$ concentrations. Pol does not incorporate a correct nucleotide into the primer with an incorrect nucleotide at the end; instead, it preferentially excises the incorrect nucleotide at the 3' end of the primer. Pol has DNA polymerase activity at pHs 6.5 and 7.5 but little at pHs 5.5, 8.5, and 9.5. It has exonuclease activity at pHs 6.5, 7.5, and 8.5 but little at pHs 4.5, 5.5, and 9.5. The finding that Pol has exonuclease activity but not DNA polymerase at pH 8.5 suggests that DNA binds to Pol, but deoxynucleotide binding or incorporation does not occur at pH 8.5.

• 한국환경성돌연변이발암원학회지
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• 제9권1호
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• pp.33-46
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• 1989

본 연구는 알킬화제를 처리한 CHO-K1 세포에서 DNA 복제억제와 그 회복과정의 분자론적 기작을 규명할 목적으로 방사선 이중 표지에 의한 DNA 합성율의 측정, 알칼리 자당 농도구배 초원심분리법에 의한 DNA 분자량과 후복제 회복율을 측정하여 다음과 같은 결과를 얻었다. (1) 1mM methyl methanesulfonate (MMS)와 1nM N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) 이하의 낮은 농도의 처리군에서는 DNA 합성율이 급격히 감소하였으나, 2 mM MMS, 2mM MNNG이상의 농도에서는 그 감소양상이 둔화되었다, (2) DNA 합성율은 알킬화제의 처리 직후 감소하였다가 시간경과에 따라 회복되어 처리후 4시간 째에는 대조군 수준 또는 그 이상으로 회복되었다.