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Xue Ying Tao,Jae Young Choi,Woo Jin Kim,Qin Liu,Song Eun Kim,Saes Byeol An,Seok Hee Lee,Soo Dong Woo,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.04
ORF78 (ac78) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a baculovirus core gene of unknown function. To determine the role of ac78 in baculovirus life cycle, an ac78-deleted mutant AcMNPV, Ac78KO, was constructed. Quantitative PCR analysis revealed that ac78 is a late gene in the viral life cycle. After transfection into Spodoptera frugiperda cells, Ac78KO produced a single-cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was also confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Analysis of BV and occlusion derived virus (ODV) revealed that AC78 is associated with both forms of the virions and is a structural protein located to viral envelope. Electron microscopy showed that ac78 also plays an important role in embedding of ODV into occlusion body. This study therefore demonstrates that AC78 is a late virion associated protein and is essential for the viral life cycle.
Xue Ying Tao,Jong Yul Roh,Jae Young Choi,Yong Wang,Qin Liu,Soo Dong Woo,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05
Autographa californica nucleopolyhedrovirus (AcMNPV) has a large doublestrand DNA genome of approximately 134 kbp and comprises 156 open reading frames (ORFs). To elucidate DNA replication cascade of AcMNPV, we developed a novel baculovirus genome that can be maintained in Escherichia coli as a plasmid and can infect susceptible lepidopteran insect cells. This genome, named bAc-MK, contains a mini-F replicon and a kanamycin resistance marker. Using a convenient Tn7 transposon-based system, pPCS-S, which contains an ampicillin resistance gene, 56 single ORF-truncated mutants were generated by random insertion into bAc-MK genome. These single ORF-truncated mutants were independently transfected into Sf9 cells to verify viral replication. Interestingly, both lef-1 and p48 knockout mutants showed normal viral replication in infected cells, which are reported to essential for viral replication. These results suggest that these single ORF-truncated mutants are useful for elucidation of viral replication cascade.
Xue Ying Tao,Jae Young Choi,Woo Jin Kim,Qin Liu,Song Eun Kim,Saes Byeol An,Seok Hee Lee,Soo Dong Woo,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.04
ORF11 (ac11) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a highly conserved gene of unknown function. To determine the role of ac11 in baculovirus life cycle, an ac11-knockout mutant AcMNPV, Ac11KO, was constructed. qPCR analysis revealed that ac11 is an early gene in the life cycle. After transfection into Spodoptera frugiperda cells, Ac11KO produced a single cell infection phenotype indicating that no infectious budded viruses (BVs) were produced. The defection in BV production was confirmed by both viral titration and Western blot. However, viral DNA replication is unaffected. Electron microscopy showed that ac11 is required for nucleocapsids envelopment to form ODV and their subsequent embedding into OB. This study therefore demonstrates that ac11 is an early gene which is essential for the viral life cycle.
Functional analysis of Autographa californica multiple nucleopolyhedrovirus ac78 and ac79
Xue Ying Tao,Jae Young Choi,Jae Su Kim,Qin Liu,Jong Bin Park,Joo Hyun Lee,Soo Dong Woo,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.10
Among 154 putative ORFs of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), ac78 and ac79 are highly conserved genes in baculovirus, but their functions in the virus life cycle have been unknown so far. To determine their roles in AcMNPV replication, knockout mutants, ac78KO and ac79KO, were constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that both of ac78 and ac79 transcripts were first detected at 6 hours post-infection, and accumulated to maximum at 24 hours post-infection, suggesting that both of ac78 and ac79 are belong to late gene. When the genomic DNA of ac78KO was transfected into Sf9 cells, viral replication was restricted to a single cell infection. These results demonstrated that the ac78 play an important role in BV production, and therefore is essential for AcMNPV to mount a successful infection. Whereas Sf9 cells infected with the ac79KO showed normal viral symptoms such as rounding and swelling, OBs were not observed from majority of infected cells. These results suggested that the ac79 might play an important role in OB production.
신혈관성 고혈압 흰쥐 선장에서 심방 이뇨 호르몬 발현 변화
김수완(Soo Wan Kim),이종은(Jong Un Lee),오윤화(Yoon Wha Oh),이영순(Ying Shun Li),김남호(Nam Ho Kim),최기철(Ki Chul Choi) 대한신장학회 2002 Kidney Research and Clinical Practice Vol.21 No.5
배 경 : Two-kidney, one clip (2K1C) 신혈관성 고혈압 신장에서 심방 이뇨 호르몬 (atrial natriuretic peptide, ANP) 계의 변화를 알아보고자 하였다. 방 법 : 실험동물은 Sprague-Dawley 숫쥐를 사용하였다. 실험군은 왼쪽 신동맥에 클립을 끼웠으며, 대조군은 다른 절차는 실험군과 같으나 신동맥에 클립을 끼우지 않았다. 4주 후 혈압을 측정한 후 클립 신장 및 반대쪽 신장을 적출하여 조직 ANP, natriuretic peptide receptor-A (NPR-A) 및 natriuretic peptide recepr-C (NPR-C) mRNA 발현을 역전사 중합효소 연쇄반응으로 조사하였다. 결 과 : 수축기 혈압은 실험군에서 대조군에 비하여 유의하게 높았다. 이와 동시에 ANP mRNA 발현은 클립 신장에서 유의하게 감소하였으며, 반대편 신장에서 증가하였다 NPR-A 및 NPR-C 발현은 클립 신장에서 감소하고 반대쪽 신장에서 증가하였다. 결 론 : 신혈관성 고혈압 신장의 ANP와 NPR 발현이 클립 쪽과 반대 쪽에서 각각 다르게 조절되리라 생각된다. Background : The present study was aimed to determine the pathophysiological implications of local atrial natriuretic peptide (ANP) system in the kidney in twokidney, one clip (2K1C) hypertension. Methods : Rats were made 2K1C hypertensive, and their mRNA expressions of ANP and natriuretic peptide receptors (NPR) were determined in the clipped and contralateral kidneys by reverse transcription-polymerase chain reaction. Results : The expression of ANP was decreased in the clipped kidney and increased in the contralateral kidney. Similarly, the expression of both NPR-A and NPR-C was decreased in the clipped kidney and increased in the contralateral kidney. Conclusion : These findings indicate a differentially-altered ANP system in the clipped and the contralateral kidneys in 2K1C hypertension.
Chlorpropamide 처리 흰쥐 신장에서 Aquaporin - 2 단백 발현 감소
김수완(Soo Wan Kim),이종은(Jong Un Lee),이영순(Ying Shun Li),오윤화(Yoon Wha Oh),김남호(Nam Ho Kim),최기철(Ki Chul Choi) 대한신장학회 2002 Kidney Research and Clinical Practice Vol.21 No.3
배 경 : 신장의 aquaporin(AQP) 수분 통로 발현이 혈당 변화에 의해 영향 받는지 알아보고자 하였다. 실험방법: 혈당강하제인 chlorpropamide(40 ㎎/100 g)를 경구 투여하고 나서 7일 뒤 신장에서 AQP1-3 수분통로 단백 발현을 Western blot 분석에 의해 조사하였다. AQP2 이동 기전의 변조 여부는 조직의 막 분획과 세포질 분획에서 각각 AQP2 발현을 분석하여 결정하였다. 나아가서 AQP 통로 발현 변조의 상위 기전을 알아보고자 adenylyl cyclase type Ⅵ 단백 발현도 Western blot 분석에 의해 조사하였다. 결 과 : 혈당은 chlorpropamide 투여군에서 대조군에 비하여 유의하게 감소하였다(64±8 및 106±7 ㎎/dL, 각 6례, p<0.01). 동시에 AQP2 단백 발현은 신장의 피질, 외수질, 내수질에서 모두 유의하게 감소하였다. Chlorpropamide 투여군에서 막 분획의 AQP2 단백 발현은 세포질 분획의 AQP2 단백 발현과 평행하게 감소하여 이동 기전에 변조가 없음을 시사하였다. AQP1과 AQP3 단백 발현은 유의한 변화를 보이지 않았다. Type Ⅵ adenylyl cyclase 단백 발현은 유의한 변화를 보이지 않았다. 결 론 : 이상의 실험성적은 저혈당에 의해 신장내 AQP2 단백 발현이 감소 조절됨을 시사하였다. Background : Whether blood glucose levels may change the regulation of aquaporin(AQP) water channels in the kidney was investigated. Methods : Male Sprague-Dawley rats were treated with chlorpropamide(40 mg/100 g body weight per day, per oral, for 7 days), and their expression of AQP1-3 and type Ⅵ adenylyl cyclase proteins was determined in the kidney. Results : Following the treatment with chlorpropamide, the blood glucose level was significantly decreased compared with that in the control(64±8 vs 106±7 ㎎/dL, n=6 each, p<0.01). Accordingly, the expression of AQP2 proteins was decreased in the cortex, outer medulla, and inner medulla. The AQP2 targeting was not significantly altered, as evidenced by parallel decreases of its expression in the membrane and the cytoplasmic fractions. No significant changes were observed in the expression of either AQP1 or of AQP3. The protein expression of type Ⅵ adenylyl cyclase was not significantly altered. Conclusion : These results suggest that hypoglycemia attenuates the expression of AQP2 water channels in the kidney.
Xue Ying Tao,Jong Yul Roh,Jae Young Choi,Yong Wang,Qin Liu,Jong Bin Park,Soo Dong Woo,Byung Rae Jin,Yeon Ho Je 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10
Autographa californica nucleopolyhedrovirus (AcMNPV) has a large doublestrand DNA genome of approximately 134 kbp and harbors 156 open reading frames (ORFs). To elucidate DNA replication cascade of AcMNPV, we developed a novel baculovirus genome that can be maintained in Escherichia coli as a plasmid and can infect susceptible lepidopteran insect cells. This genome, named bAc-MK, contains a mini-F replicon and a kanamycin resistance marker. Using a convenient Tn7 transposon-based system, pPCS-S, 55 single ORF-truncated mutants were generated by random insertion into bAc-MK genome. These single ORF-truncated mutants were independently transfected into Sf9 cells, 16 of them were found affecting viral replication since they defected in producing polyhedra. Furthermore, to verify the pathogenicity of the single ORF-truncated mutants, the remaining 39 mutants were subjected to bioassay to Spodoptera exigua 3rd instar larvae. Among them, ac9-, ac49-, ac103- and ac105-knockout mutants showed higher mortality compared to that of bAc-MK. These results suggested that these ORFs could be related to pathogenicity of AcMNPV.
( Jin Ho Choi ),( Shu Ying Zhang ),( Kyung Woo Park ),( Young Seok Cho ),( Byung Hee Oh ),( Myoung Mook Lee ),( Young Bae Park ),( Hyo Soo Kim ) 생화학분자생물학회 2005 BMB Reports Vol.38 No.2
5-HT2A is one of major serotonin receptor that is involved in the action of serotonin-targeting drugs. Previous clinical studies have shown an unexpected association between lower cholesterol level and psychiatric diseases, in which T102C polymorphism of HTR2A, gene of 5-HT2A serotonin receptor, might be involved. Therefore, we hypothesized a potential association between lower cholesterol level and T102C polymorphism. The effect of the T102C polymorphism on the serum lipid profiles of 646 subjects without specific psychiatric disease was investigated. Genotype was determined by polymerase chain reaction and restriction fragment length polymorphism analysis. There were significantly lower levels of total cholesterol (193.6±35.0 versus 202.1±45.5 mg/dl, p = 0.016) and HDL-cholesterol (42.7 ±11.6 versus 46.3±12.7 mg/dl, p = 0.004) in CC genotype than non-CC genotypes. Moreover, multivariate analysis showed that the CC genotype is a strong predictor of a lower HDL-cholesterol level (p < 0.001). In conclusion, this study shows that the CC genotype of the HTR2A gene is related to lower HDL-cholesterol level in Koreans. This is the first demonstration showing the potential genetic relationship between the serotonin receptor gene polymorphism and the HDL-cholesterol level.
Choi, Jin-Ho,Zhang, Shu-Ying,Park, Kyung-Woo,Cho, Young-Seok,Oh, Byung-Hee,Lee, Myoung-Mook,Park, Young-Bae,Kim, Hyo-Soo Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.2
5-HT2A is one of major serotonin receptor that is involved in the action of serotonin-targeting drugs. Previous clinical studies have shown an unexpected association between lower cholesterol level and psychiatric diseases, in which T102C polymorphism of HTR2A, gene of 5-HT2A serotonin receptor, might be involved. Therefore, we hypothesized a potential association between lower cholesterol level and T102C polymorphism. The effect of the T102C polymorphism on the serum lipid profiles of 646 subjects without specific psychiatric disease was investigated. Genotype was determined by polymerase chain reaction and restriction fragment length polymorphism analysis. There were significantly lower levels of total cholesterol ($193.6{\pm}35.0$ versus $202.1{\pm}45.5\;mg/dl$, p = 0.016) and HDL-cholesterol ($42.7{\pm}11.6$ versus $46.3{\pm}12.7\;mg/dl$, p = 0.004) in CC genotype than non-CC genotypes. Moreover, multivariate analysis showed that the CC genotype is a strong predictor of a lower HDL-cholesterol level (p < 0.001). In conclusion, this study shows that the CC genotype of the HTR2A gene is related to lower HDL-cholesterol level in Koreans. This is the first demonstration showing the potential genetic relationship between the serotonin receptor gene polymorphism and the HDL-cholesterol level.