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( Kwan Ho Park ),( Young Cheol Cho ),( Seong Hee Nam ),( Jae Sam Hwang ),( Si Kab Nho ) 한국잠사학회 2012 International Journal of Industrial Entomology Vol.25 No.2
Serine proteases are major insect enzymes involved in the digestion of dietary proteins and in the process of blood meal digestion. In this study, cDNA was constructed using the whole body of Laccotrephes japonensis. The flanking sequences of the 5- and 3- end of this gene were characterized by RACE-PCR. Sequence analysis showed that this gene contained a 963-bp ORF encoding 320 amino acids. The deduced amino acid sequence showed 62% identity with the Creontiades dilutus serine protease, 58% with the Lygus lineolaris trypsin precursor, and 54% with the Triatoma infestans salivary trypsin. To assess the expression of the L. japonensis serine protease (JGsp), the JGsp gene was cloned into a baculovirus transfer vector, pBac-1, and expressed in Sf9 cells (Spodoptera frugiperda). SDS-PAGE and western blot analysis have shown that the JGsp recombinant protein was a monomer with a molecular weight of about 32 kDa. Recombinant JGsp has shown activity in the protease enzyme assay using gelatin as a substrate.
Kwan-Ho Park,Young-Cheol Choi,Jong-Gil Kim,Ji-Young Choi,Won-Tae Kim,Si-Kab Nho,Jae-Sam Hwang 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05
Protease from various sources have been studied biotecnologically. For biotechnological applications, one highly preferred enzyme is protease. There have been no reports of cloned genes encoding digestive proteases in the Laccotrephes japonenis, Ranatra unicolor, Muljarus japonicus. These insects are considered to be a predator of aquatic insects. RT-PCR was used to amplify cDNA fragments for digestive proteases from total RNA the hole body of the insects. The flanking sequences of the 5'- and 3'- end of the these genes were characterized by RACE-PCR. Sequence analysis showed that these genes contained complete ORF. The deduced amino acid sequences of these protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin-like serine proteinase, 54% to Triatonatoma infestans salivary trypsin. To generate Laccotrephes japonensis serine protease, the DNA fragment coding for serine protease is cloning into suttle vector pBACⅠ, named pBAC1-JG and infected to Spodoptera frugiperda (sf9) insect cell. The cDNA encoding JG was expressed as a 32-kDa polypeptide in baculovirus infected insect cells and the recombinant protein showed activity in the protease enzyme assay using gelatin as a substrate.
Yong-Duck Kim,Hak-Gon Kim,Jong-Cheol Kim,Seon Jeong Sim,Ji-Yun Min,Jung-Gyu Hwang,Seung Mi Kang,Hyun-Shik Moon,Jong-Kab Kim,Myung-Suk Choi 경상대학교 농업생명과학연구원 2013 농업생명과학연구 Vol.47 No.1
The influence of the culture media on the growth and catechins and caffeine production was investigated in adventitious root cultures of tea tree. The growth rate of adventitious roots was higher than that of solid medium. Growth rate of adventitious roots was noted to be optimal in N6 liquid medium. Yields of (-)-epicatechin gallate (ECG) and caffeine were the highest when adventitious root cultures were maintained in Nitsch and B5 liquid medium, respectively. The production of (-)-epicatechin (EC) and (-)-epigallocatechin gallate (EGCG) from adventitious roots was maximal when cultured in 1/2MS liquid medium. The adventitious root extracts of tea tree produced on catechins as EC (20.77 mg/g) and EGCG (1.84 mg/g) in 1/2MS medium; EGC (24.39 mg/g) and caffeine (12.97 mg/g) in B5 medium and (-)-epigallocatechin (ECG) (2.16 mg/g) in Nitsch liquid medium.
Park, Kwan Ho,Choi, Young Cheol,Nam, Seong Hee,Hwang, Jae Sam,Nho, Si Kab Korean Society of Sericultural Science 2012 International Journal of Industrial Entomology Vol.25 No.2
Serine proteases are major insect enzymes involved in the digestion of dietary proteins and in the process of blood meal digestion. In this study, cDNA was constructed using the whole body of Laccotrephes japonensis. The flanking sequences of the 5- and 3- end of this gene were characterized by RACE-PCR. Sequence analysis showed that this gene contained a 963-bp ORF encoding 320 amino acids. The deduced amino acid sequence showed 62% identity with the Creontiades dilutus serine protease, 58% with the Lygus lineolaris trypsin precursor, and 54% with the Triatoma infestans salivary trypsin. To assess the expression of the L. japonensis serine protease (JGsp), the JGsp gene was cloned into a baculovirus transfer vector, pBac-1, and expressed in Sf9 cells (Spodoptera frugiperda). SDS-PAGE and western blot analysis have shown that the JGsp recombinant protein was a monomer with a molecular weight of about 32 kDa. Recombinant JGsp has shown activity in the protease enzyme assay using gelatin as a substrate.
Thermal Impact Characteristics by Forest Fire on Porcelain Insulators for Transmission Lines
Lee, Won-Kyo,Choi, In-Hyuk,Choi, Jong-Kee,Hwang, Kab-Cheol,Han, Se-Won The Korean Institute of Electrical and Electronic 2008 Transactions on Electrical and Electronic Material Vol.9 No.4
In this study the thermal impact characteristics by forest fire are extensively investigated using temperature controlled ovens. The test conditions for thermal impact damage are simulated according to the characteristics of natural forest fire. The test pieces are suspension porcelain insulators made by KRI in 2005 for transmission lines. In the thermal impact cycle tests with $300\;^{\circ}C$ thermal impact gradient (-70 to $230\;^{\circ}C$), cycling in 10 minute periods, no critical failures occurred in the test samples even with long cycle times. But in tests with thermal impact gradient from room temperature to $200-600\;^{\circ}C$, cycling in 10 to 30 minute periods, there were critical failures of the porcelain insulators according to the thermal impact gradient and quenching method. In the case of thermal impact by forest fire, it was found of that duration time is more important than the cycling time, and the initiation temperature of porcelain insulator failures is about $300\;^{\circ}C$, in the case of water quenching, many cracks and fracture of the porcelain occurred. It was found that the thermal impact failure is closely related to the displacement in the cement by thermal stress as confirmed by simulation. It was estimated that the initiation displacement by the thermal impact of $300\;^{\circ}C$ is about 0.1 %. Above 1% displacement, it is expected that the most porcelain insulators would fail.