• BMB Reports
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• 제40권1호
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• pp.107-112
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• 2007

To study the functioning of HSP70 in Escherichia coli, we selected NtHSP70-2 (AY372070) from among three genomic clones isolated in Nicotiana tabacum. Recombinant NtHSP70-2, containing a hexahistidine tag at the amino-terminus, was constructed, expressed in E. coli, and purified by $Ni^{2+}$ affinity chromatography and Q Sepharose Fast Flow anion exchange chromatography. The expressed fusion protein, $H_6NtHSP70$-2 (hexahistidine-tagged Nicotiana tabacum heat shock protein 70-2), maintained the stability of E. coli proteins up to 90$^{\circ}C$. Measuring the light scattering of luciferase (luc) revealed that NtHSP70-2 prevents the aggregation of luc without ATP during high-temperature stress. In a functional bioassay (1 h at 50$^{\circ}C$) for recombinant $H_6NtHSP70$-2, E. coli cells overexpressing $H_6NtHSP70$-2 survived about seven times longer than those lacking $H_6NtHSP70$-2. After 2 h at 50$^{\circ}C$, only the E. coli overexpressing $H_6NtHSP70$-2 survived under such conditions. Our NtHSP70-2 bioassays, as well as in vitro studies, strongly suggest that HSP70 confers thermo-tolerance to E. coli.

• Clinical and Experimental Pediatrics
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• 제49권5호
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• pp.539-544
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• 2006

목 적 : 가와사끼병의 진단 지표로서 NT-proBNP의 유용성을 살펴보고자 하였다. 방 법 : 가와사끼병을 진단 받은 환아 58명에서 발병 10일 이내 급성기에 전기화학발광면역법(Electrochemiluminescence immunoassay)를 이용하여 NT-proBNP를 측정하였고, 그 중 51명에서 회복기(발병 60에서 81일 이내)에 NT-proBNP 값을 측정하여 비교하였다. 가와사끼병과 임상적으로 구별하기 어려운 급성 열성 질환이 있는 환아 34명을 대조군으로 하여 NT-proBNP를 측정하여 가와사끼병 환자군과 비교하였다. 또한 가와사끼병 급성기에 심장 초음파검사를 시행하여 심낭삼출과 관상동맥 병변의 여부를 확인하였고 좌심실 확장기말 내경(LVIDd)과 수축기말 내경(LVIDs), 좌심실 구출율(LVEF)과 좌심실 단축율(LVSF)을 측정하여 NT-proBNP 측정치와의 상관관계를 조사하였다. 가와사끼병과 다른 질환의 감별진단을 위해 receiveroperating characteristic curve를 이용하여 NT-proBNP의 cutoff value를 구하였다. 결 과 : 가와사끼병 급성기 NT-proBNP 측정치는 대조군에 비해 유의하게 높았다($1,501.6{\pm}2,132.6$ vs. $139.0{\pm}88.8pg/mL$, P<0.0001). 51명의 가와사끼병 환자군에서 NT-proBNP 값은 급성기에 증가하고 회복기에 크게 감소하였다($1,466.0{\pm}2,173.2$ vs. $117.5{\pm}95.5pg/mL$, P<0.0001). 가와사끼병의 감별진단을 위한 NT-proBNP의 cutoff value를 260 pg/mL으로 하였을 때 민감도 93%, 특이도 88%를 보였다. NT-proBNP는 심낭삼출이 있는 환아에서 심낭삼출이 없는 환아에 비해 높았다($1,784.2{\pm}1,903.1$ vs. $1,384.4{\pm}2,232.6pg/mL$). 관상동맥 병변이 있는 환아(n=3)가 적어 NT-proBNP와의 관계를 확인할 수 없었다. NT-proBNP 측정치와 LVEF index(r=0.104, P=0.46) 및 LVIDd index(r=0.171, P=0.22)은 상관관계가 없었다. 결 론 : NT-proBNP는 가와사끼병 급성기에 크게 증가하며 회복기에 감소한다. NT-proBNP >260 pg/mL일 때 가와사끼병을 강력히 의심할 수 있으며, NT-proBNP는 가와사끼병을 진단하는 지표로 사용될 수 있을 것으로 생각된다.

• Reproductive and Developmental Biology
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• 제28권4호
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• pp.253-256
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• 2004

Porcine fibroblasts were transferred into enucleated bovine oocytes for the interspecies nuclear transfer (NT). After NT, the embryos were cultured in three different culture systems. The media used for the experiment were CR1aa and NCSU23. The culture systems used for the experiment were: 1. Culture in CR1aa for 7 days (CR). 2. Culture in CR1aa for 2 days and subsequently in NCSU23 for 5 days (CR-NC). 3. Culture in NCSU23 for 7 days (NC). Bovine (intraspecies) NT group was used as a control. The oocytes in bovine NT group were treated the same as interspecies NT embryos except using bovine fibroblasts as nuclear donors. Regardless of their nuclear origin (interspecies vs bovine), the embryos in CR (68.4% vs 77.2%) and CR-NC (67.8% vs 70.5%) showed better developmental competence to the 2-cell stage (p<0.05) than those in NC (41.0% vs 10.0%). Bovine NT embryos in CR-NC did not develop over the 4-cell stage after the medium replacement, while interspecies NT embryos in CR-NC continued to develop and could reach over the 8-cell stage (12.2%). Blastocysts were only found in bovine NT group (17.4%), but no blastocyst was found in interspecies NT group. This study suggests that the development of interspecies NT embryos mostly depends on their recipient cytoplasm during the culture in vitro.

• Journal of Plant Biotechnology
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• 제2권2호
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• pp.79-82
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• 2000

We have isolated a cDNA encoding a calcium-dependent protein kinase (CDPK) in Nicotiana tabacum, which was designated NtCDPK1. Accumulation of the NtCDPK1 mRNA was stimulated by various stimuli, including phytohormones, CaCl$_2$ wounding, fungal elicitors, chitin and methyl jasmonate. The NtCDPK1 gene encodes a functional Ser/Thr protein kinase of which phosphorylation activity is strongly induced by calcium. By analyzing expression of the NtCDPK1-GFP fusion protein and by immunoblotting with antibody which reacts with NtCDPK1, we found that NtCDPK1 is localized in membrane and nucleus in plant cells. Silencing expression of the NtCDPK1 transgene resulted in marked decrease of lateral root development in the transgenic tobacco plants. Yeast two hybrid screening using NtCDPK1 as a bait identified a tobacco homologue of proteasome regulatory subunit 21D7, designated Nt21D7. The 21D7 mRNA has been shown to be predominantly expressed in proliferating tissues in the cell cycledependent manner in carrot. The recombinant NtCDPK1 protein associated with Nt21D7 in vitro, and could phosphorylate the Nt21D7 protein in vitro in the presence of calcium, suggesting that Nt21D7 protein is a natural substrate of NtCDPK1 in tobacco. These results suggest that NtCDPK1 may regulate tell proliferation processes, such as lateral root formation, by regulating specificity and/or activity of proteasome-mediated protein degradation pathway.

• 한국수정란이식학회지
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• 제23권2호
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• pp.101-108
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• 2008

This study investigated the developmental ability and gene expression of somatic cell nuclear transfer embryos using ear skin fibroblast cells derived from miniature pig. When miniature pig (m) and landrace pig (p) were used as donor cells, there were no differences in cleavage (79.2 vs. 78.2%) and blastocyst rates (27.4 vs. 29.7%). However, mNT blastocysts showed significantly higher apoptosis rate than that of pNT blastocysts (6.1 vs. 1.7%) (p<0.05). The number of nuclei in pNT blastosysts was significantly higher than that of mNT (35.8 vs. 29.3) (p<0.05). Blastocysts were analyzed using Realtime RT-PCR to determine the expression of Bax-${\alpha}$, Bcl-xl, H19, IGF2, IGF2r and Xist. Bax-${\alpha}$ was higher in mNT blastocyst than pNT blastocyst (p<0.05). There was no difference in Bcl-xl between two NT groups. Bax-${\alpha}$/Bcl-xl was, however, significantly higher in mNT blastocyst compared to pNT. The expression of imprinting genes were aberrant in blastocysts derived from NT compared to in vivo blastocysts. H19 and IGF2r were significantly lower in mNT blastocysts (p<0.05). The expression of IGF2 and Xist was similar in two NT groups. However, imprinting genes were expressed aberrantly in mNT compared to pNT blastocysts. The present results suggest that the NT between donor cells derived from miniature pig and recipient oocytes derived from crossbred pig might affect reprogramming of donor cell, resulting in high apoptosis and aberrant expression patterns of imprinting genes.

• 한국작물학회지
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• 제47권5호
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• pp.384-389
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• 2002

No-tillage (NT) practice for corn production has advantages of reduction of soil erosion and energy conservation. Research on effects of NT for sweet corn or super sweet corn is very limited. Hybrids of sugary (su) and shrunken-2 (sh2) were tested under NT and conventional tillage (CT) practices to investigate plant characters, ear characters, fresh yield, and grain yield. Sugary hybrids were Golden Cross Bantam 70 (GCB70), Sprint, Geumdanok, and Danok3. Shrunken-2 hybrids were BSS9472, Cambella90, GSS9299, Jubilee, KS-Y-65, and Chodangok1. Emergence rates under NT were lower than those under CT for su, while there was no difference between tillage systems for sh2. There were no differences between CT and NT for days to tasseling and silking, plant height, and ear height for both su and sh2. Ear characters such as ear length, number of kernel rows, number of kernels per row, and t100-kernel weight under NT were not significantly different from those under CT. There were no differences between two tillage practice for fresh and grain yield, rather they showed trend of increases under NT practices. Results from this trial indicate that NT practice for both su and sh2 cultivation may be possible to recommend to farmers.

• The Korean Journal of Physiology and Pharmacology
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• 제4권3호
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• pp.227-234
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• 2000

Many reports suggest that neurotensin (NT) in the gastrointestinal tract may play a possible role as a neurotransmitter, a circulating hormone, or a modulator of motor activity. NT exerts various actions in the intestine; it produces contractile and relaxant responses in intestinal smooth muscle. This study was designed to investigate the effect of NT on motility of antral circular muscle strips in guinea-pig stomach. To assess the role of $Ca^{2+}$ influx in underlying mechanism, slow waves were simultaneously recorded with spontaneous contractions using conventional intracellular microelectrode technique. At the concentration of $10^{-7}$ M, where NT showed maximum response, NT enhanced the magnitude $(863{\pm}198%,\;mean\;SEM,\;n=13)$ and the frequency $(154{\pm}10.3%,\;n=11)$ of spontaneous contractions. NT evoked a slight hyperpolarization of membrane potential, tall and steep slow waves with abortive spikes $(278{\pm}50%,\;n=4).$ These effects were not affected by atropine $(2\;{\mu}M),$ guanethidine $(2\;{\mu}M)$ and tetrodotoxin (0.2μM). NT-induced contractile responses were abolished in $Ca^{2+}-free$ solution and reduced greatly to near abolition by $10\;{\mu}M$ of verapamil or 0.2 mM of $CdCl_2.$ Verapamil attenuated the effects of NT on frequency and amplitude of the slow waves. Taken together, these results indicate that NT enhances contractility in guinea-pig gastric antral circular muscle and $Ca^{2+}$ influx through the voltage-operated $Ca^{2+}$ channel appears to play an important role in the NT-induced contractile mechanism.

• 생명과학회지
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• 제18권8호
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• pp.1023-1030
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• 2008

SIPK와 WIPK의 상위 단계 인산화 효소로 알려진 NtMEK2가 DEX 유도성 시스템에 의해 밝혀졌다. 이 NtMEK2 유전자가 지속적으로 활성화된 돌연변이체인 $NtMEK2^{DD}$의 발현은 SIPK와 WIPK를 활성화 시켜 주므로 과민감 반응과 같은 세포 괴사를 야기하는 것으로 나타나 NtMEK2-SIPK/WIPK 체계가 담배에서 방어 반응을 조절하고 있음을 알 수 있었다. 그러나 NtMEK2-SIPK/WIPK 체계에 의해서 조절 되는 하위 기질이나 방어관련 유전자들에 대한 연구는 아직 미비한 상태이다. 그래서 본 연구는 NtMEK2-SIPK/WIPK 체계에 매개되는 하위 유전자들을 분리하기 위하여 $NtMEK2^{DD}$ 형질전환 식물체를 이용해 ACP에 기초한 DDRT-PCR을 수행하였다. 그 결과 본 연구를 통해 처음으로 pI2-4, MTS2, SINA, CDM1, HRGP 및 DEG45를 포함해 여섯 개의 DEG들을 선발하였다. 이 유전자들의 발현은 $NtMEK2^{DD}$ 형질전환에서 다시 확인하였으며 특히 pI2-4, CDM1, HRGP의 유전자 발현은 다른 유전자들과 비교해 볼 때 살리실산과 담배모자이크바이러스에 강하게 반응하여 증폭됨을 알 수 있었다. 이러한 결과를 볼 때 NtMEK2-SIPK/WIPK 체계에 의해 조절되는 세 개의 유전자는 병저항성에 관여하고 있음을 제시한다 하겠다.

• 한국임상수의학회지
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• 제35권2호
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• pp.35-38
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• 2018

Recently assessment of suppression of tumorigenicity 2 (ST2) level has become a useful cardiac biomarker in human medicine. This study compared serum ST2 levels and N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels between healthy dogs and dogs with chronic mitral valve disease. Twenty client-owned dogs were investigated. Dogs were divided into normal, asymptomatic, and symptomatic groups. Serum samples were used to measure levels of NT-proBNP and ST2. Samples for NT-proBNP were sent to IDEXX laboratory for analysis while ST2 levels were measured by using a canine interleukin 33 receptor ELISA kit. There was a significant difference in NT-proBNP levels between asymptomatic and symptomatic groups (P < 0.01), and between normal and symptomatic groups (P < 0.01). In contrast, ST2 levels were not relatively different between asymptomatic and symptomatic groups (P > 0.05). There was no significant difference was observed among all groups in ST2 study. The usefulness of measuring NT-proBNP level as a cardiac biomarker in dogs with chronic mitral valve disease was confirmed, but usefulness of the ST2 level was not observed. Further investigations are needed to evaluate the potential usefulness of ST2 level as a cardiac biomarker in canines.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.515-523
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• 2001

The dinoflagellates have some primitive nuclear features and are evolutionarily intermediate between prokaryotes and eukaryotes. The small subunit ribosomal RAN gene, the 5.8S ribosomal RNA gene, and the internal transcribed spacer (ITS) of Gonyaulax polyedra were cloned, and their sequences were analyzed to better understand their evolutionary position. The small subunit ribosomal RNA gene was 1,794 nt long, the large subunit ribosomal RNA gene was approximately 3,500 nt long, and the 5.8S ribosomal RNA gene was 159 nt long. The first internal transcribed spacer (ITS1) was 191 nt long, and the second internal transcribed spacer (ITS2) was 185 nt long. The intergenic spacer of the ribosomal RNA gene (IGS) was about 2,200 nt long, indicating that 5,800 nt of transcribed sequences were separated by roughly 2,200 nt of intergenic spacer. The ribosomal RNA genes were repeated many times and arranged in a head-to-tail, tandemly repeated manner. The repeating unit of ribosomal RNA gene of G. polyedra was proposed to be 8,000 nt long. Based on the lengths of ribosomal RNA, sequence alignments with representative organisms, and phylogenetic analysis on ribosomal RNA, G. polyedra appears to be one of the alveolates branched from the eukaryotic crown and, among dinoflagellates, it seems to not have emerged early.