• 생명과학회지
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• 제15권5호
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• pp.755-762
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• 2005

세포 내에서 매우 적은 양으로 존재하는 RNA 전사체와 기능적으로 동일하거나 비슷한 RNA를 RNA 중합효소를 써서 in vitro에서 생화학적으로 의미 있는 양 만큼을 합성할 수 있다. T7 RNA중합효소를 발현하는 재조합 유전자를 지닌 대장균주 BL21/pAR1219로부터 순수한 T7 RNA중합효소를 손쉽게 얻는 방법을 본 논문에서 소개한다. 황산암모늄 분획화와 sephadex SP 컬럼 크로마토그래피법으로써 여타의 방법과 비교하여 더 간단하고 빠르게, 그리고 경제적으로 T7 RNA 중합효소를 분리할 수 있었다. 정제된 T7 RNA중합효소를 이용하여 보통의 화학적 합성법으로 불가능한 긴 길이(1.54 kb)의 RNA전사체를 합성 하였다. 한편,정제된 T7 RNA중합효소에 의해 생성된 망치머리 리보자임은 표적 RNA를 in vitro에서 절단함으로써, 생성된 RNA가 생화학적 기능성을 유지한다는 것을 입증하였다 따라서 본 연구에서 소개되는 절차들은 다양한 길이의 RNA를 목적에 따라 간단하고 경제적으로 합성하는데 유용하게 이용될 수 있다.

• 한국자기공명학회:학술대회논문집
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• 한국자기공명학회 2002년도 International Symposium on Magnetic Resonance
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• pp.67-67
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• 2002

• BMB Reports
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• 제39권3호
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• pp.329-334
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• 2006

RNAi (RNA interference) has become a popular means of knocking down a specific gene in vivo. The most common approach involves the use of chemically synthesized short interfering RNAs (siRNAs), which are relatively easy and fast to use, but which are costly and have only transient effects. These limitations can be overcome by using short hairpin RNA (shRNA) expression vectors. However, current methods of generating shRNA expression vectors require either the synthesis of long (50-70 nt) costly oligonucleotides or multi-step processes. To overcome this drawback, we have developed a one-step short-oligonucleotides-based method with preparation costs of only 15% of those of the conventional methods used to obtain essentially the same DNA fragment encoding shRNA. Sequences containing 19 bases homologous to target genes were synthesized as 17- and 31-nt DNA oligonucleotides and used to construct shRNA expression vectors. Using these plasmids, we were able to effectively silence target genes. Because our method relies on the onestep ligation of short oligonucleotides, it is simple, less error-prone, and economical.

• 한국환경보건학회:학술대회논문집
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• 한국환경보건학회 2005년도 가을학술대회
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• pp.166-168
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• 2005

The induction of heme oxygenase-1(HO-1) by ultraviolet(UV) radiation provides a protective defense against oxidative stress, and has been well demonstrated in UVA-irradiated skin, but not UVB. In this study in mice, we show that the UVB(290-320nm) radiation can be attributed to the induction of cutaneous heme oxygenase-1. The expression of HO-1 mRNA was assessed in vivo by the reverse transcription-polymerase chain reaction (RT-PCR) analysis, and HO-1 enzyme activity was measured in microsomal preparation from irradiated mice. The mRNA level of HO-1 increases in liver and skin from 24h to 72h after UVB($3KJ/m^3$) radiation. The results of gene expression were same pattern of HO enzyme activity in skin, but not in liver. HO-1 mRNA in liver resulted in a progressive increase to 96h after UVB radiation, but HO activity in liver increased to 48h. This finding indicates that UVB radiation is an important inducer of HO-1 and increases in HO activity may protect tissues directly or indirectly from oxidative stress.

• 한국환경보건학회지
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• 제34권1호
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• pp.49-54
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• 2008

The induction of heme oxygenase-1(HO-1) by UV radiation provides a protective defence against oxidative stress, and has been well demonstrated in skin irradiated with UVA, but not UVB. In this study, we show that the induction of cutaneous HO-l can be attributed to UVB radiation. The expression of HO-1 mRNA was assessed in vivo by reverse transcription-polymerase chain reaction (RT-PCR) analysis, and HO-1 enzyme activity was measured in microsomal preparation from irradiated mice. The mRNA level of HO-1 increases in liver and skin from 1d to 3d after UVB $(3KJ/m^2)$ exposure. The results of gene expression were same pattern of HO-1 enzyme activity in skin, but not in liver. HO-1 mRNA in liver resulted in a progressive increase to 4d after UVB exposure, but HO-1 activity in liver increased to 2d. This finding indicates that UVB radiation is an important inducer of HO-1 and increases in HO activity may protect tissue directly or indirectly from oxidative stress.

• Bulletin of the Korean Chemical Society
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• 제32권10호
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• pp.3767-3769
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• 2011

• 생명과학회지
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• 제19권12호
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• pp.1808-1814
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• 2009

방사선에 대한 방호와 면역기능 조절을 목적으로 새로운 생약복합조성물인 HemoHIM을 개발하였다. 식품 원료로 사용 가능한 생약재 3종 당귀, 천궁, 백작약의 에탄올 분획을 열수추출물에 첨가하여 HemoHIM을 제조하였다. HemoHIM의 항알레르기 효과를 검증하기 위하여 사람 비만세포주 HMC-1을 사용해 compound 48/80으로 유도되는 히스타민 분비량과 PMA/A23187로 유도되는 염증성 사이토카인의 분비량을 측정하였다. 히스타민의 양은 형광분석법으로, 염증성 사이토카인 IL-6, IL-8, TNF-$\alpha$, GM-CSF의 양은 효소결합 면역측정법으로 측정하였다. 저농도의 HemoHIM에 의해 히스타민 분비량이 억제되었고, 모든 농도에서 IL-6, TNF-$\alpha$, GM-CSF의 분비량은 억제되었지만 IL-8은 고농도에서만 억제되었다. 사이토카인의 mRNA 발현량은 HemoHIM의 농도 의존적으로 억제되었다. 그리고 c-kit와 Fc$\varepsilon$RI의 mRNA 발현량도 모든 농도에서 억제되었지만, tryptase의 mRNA 발현량은 저 농도에서만 억제되었다. 이상의 결과로 HemoHIM이 비만세포의 활성을 억제하는 효과가 있다는 것을 알 수 있었으며, 상대적으로 독성이 적은 항알레르기 제재로 개발할 가능성을 제시한 것으로 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권1호
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• pp.97-102
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• 2008

Four hundred and forty 1-day-old Arbor Acre broilers were fed commercial starter diets with 0, 250, 750 and 2,250 mg/kg of an alpha-amylase preparation from 1 to 21 days of age to investigate the effects of an exogenous enzyme on growth performance, activities of digestive enzymes in the pancreas and anterior intestinal contents and pancreatic amylase mRNA expression. Body weight gain (BWG) and average daily gain (ADG) increased linearly (p<0.01) with increasing levels of supplementary amylase but feed conversion ratio (FCR) was not affected. There was a negative quadratic change of protease and amylase in the small intestinal contents with the increase of supplementary amylase level. The activity of intestinal trypsin was also increased (p<0.05). Lipase was unaffected by amylase supplementation (p>0.05). The pancreatic protease, trypsin, and lipase were not affected by exogenous amylase levels. Consistent with the tendency for a linear depression of amylase activity, pancreatic ${\alpha}$-amylase mRNA was down-regulated by dietary amylase supplementation. The present study suggested that oral administration of exogenous amylase affected activities of intestinal enzymes and the production of pancreatic digestive enzymes in a dose-dependent manner.

• 대한본초학회지
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• 제33권2호
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• pp.69-77
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• 2018

Objectives : Daesiho-tang (DSHT) has been widely used in the treatment of cerebral infarct in traditional medicine. However, there was not report on the anti-obesity-related diseases efficacy of DSHT. In this study, we investigated the effects for the new formulation of DSHT, on the adipocyte differentiation cycle in 3T3-L1 cells. Methods : 3T3-L1 cells were treated with DSHT (50, 100, $200{\mu}g/m{\ell}$) during differentiation for 6 days. Also, the inhibitory effect of DSHT against 3T3-L1 adipogenesis was evaluated in various stage of adipogenesis such as early (0-2day), intermediate (2-4day), and terminal stage (4-6day). The accumulation of lipid droplets was determined by Oil Red O staining. and, the expressions of genes related to adipogenesis were measured by RT-PCR and Western blot analyses. Results : DSHT showed inhibitory activity on adipocyte differentiation at 3T3-L1 preadipocytes without affect cell toxicity as assessed by measuring fat accumulation and adipogenesis. In addition, DSHT significantly reduced the expression levels of several adipocyte marker genes including proliferator activated $receptor-{\gamma}$ ($PPAR-{\gamma}$) and CCAAT/ enhancer-binding $protein-{\alpha}$ ($C/EBP-{\alpha}$). Also, the anti-adipogenic effect of DSHT was strongly limited in the intermediate (2-4 day), terminal stage (4-6 day) of 3T3-L1 adipogenesis. In addition, the DSHT treatment down- regulated mRNA expression levels of $PPAR-{\gamma}$,, $C/EBP-{\alpha}$ in mature 3T3-L1 adipocytes. Conclusions : These results suggest that, the ability of DSHT has inhibited overall adipogenesis and lipid accumulation in the 3T3-L1 cells. The new formulation of DSHT may be a promising medicine for the treatment of obesity and related metabolic disorders.

• 한국수정란이식학회지
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• 제26권3호
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• pp.209-214
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• 2011

임신의 성립 및 유지에 중요한 자궁 내막과 호르몬의 변화는 생식기관에서 발현되는 K$_{2P}$ 통로의 발현을 변화시킬 수 있다. 본 연구는 한우의 임신 자궁 내막에서 K$_{2P}$ 통로의 발현 변화가 나타나는지 그리고 프로게스테론에 의해 그 발현량이 변화되는지를 확인하고자 수행하였다. 역전사중합효소 중합반응과 웨스턴블닷 분석을 통하여 임신한 한우의 자궁 내막에서 mRNA와 단백질의 발현 변화를 조사하였다. TREK-1을 제외한 K$_{2P}$ 통로의 mRNA 발현량이 임신 자궁 내막에서 변화되었다. mRNA가 크게 변화되는 TASK-3, TREK-2, TRAAK 및 TRESK의 단백발현 변화량을 임신 자궁 내막에서 확인하였는데, TREK-2와 TRESK만 mRNA 발현 변화 양상과 동일하게 임신 자궁 내막에서 각각 7.9배, 2배 증가하였다. 자궁 내막세포에 프로게스테론(10 ${\mu}g$/mL)을 처리하였을 때 TREK-2와 TRESK는 자궁 내막 조직에서 보여준 결과와 유사하게 단백 발현량이 각각 10배, 6배 증가하였다. 이상의 결과로부터 K$_{2P}$ 통로, 특히 TREK-2와 TRESK는 프로게스테론 변화에 의해 임신 자궁 내막에서 발현량이 증가할 것으로 생각된다. 그리고 증가된 TREK-2와 TRESK는 임신에 의해 유발되는 생리학적 변화를 조절하는데 기여할 것으로 생각된다.