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Jong Yul Roh,Qin Liu,Jae Young Choi,Yong Wang,Xueing Tao,Jong Bin Park,Hee Jin Shim,Gyung Ja Choi,Jin-Cheol Kim,Yeon Ho Je 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05
A new Bacillus thuringiensis isolate 19-22 (Bt 19-22) exhibited high anti-fungal activity against barley powdery mildew (Blumeria graminis f. sp. hordei). The cry gene content of Bt 19-22 comprised cry1Aa, cry1Ab, cry1Ac and cry1D which have high insecticidal activity against lepidopteran larvae. We tried to confer a dipteran insecticidal activity to Bt 19-22 for constructing a recombinant strain which has multiple functions, anti-fungal and dual insecticidal activity. The insecticidal cry11Aa gene of B. thuringiensis was constructed under cry1Ac promoter in an E. coli-B. thuringiensis shuttle vector (pPro11A). The plasmid, pPro11A was introduced into Bt 19-22 isolate by electroporation and four transformants which had different cry gene contents were identified by PCR with cry11Aa and cry1-type specific primers. Among them, a Bt 19-22 transformant (11A/19-22 No. 7) expressed Cry11A protein (approximately 70 kDa) successfully without change of its inherent characteristics such as Cry protein expression and antifungal activity. The insecticidal activity of 11A/19-22 No. 7 was checked against Plutella xylostella and Culex pipiens. These results suggests that the recombinant strain shows dual insecticidal activity against lepidopteran and dipteran larvae as well as antifungal activity.
Fast and efficient generation of recombinant baculoviruses by in vitro transposition
Choi, Jae Young,Kim, Yang-Su,Wang, Yong,Tao, Xue Ying,Liu, Qin,Roh, Jong Yul,Woo, Soo Dong,Jin, Byung Rae,Je, Yeon Ho Springer-Verlag 2012 Applied microbiology and biotechnology Vol.96 No.5
<P>A novel recombinant bacmid, bEasyBac, that enables the easy and fast generation of pure recombinant baculovirus without any purification step was constructed. In bEasyBac, attR recombination sites were introduced to facilitate the generation of a recombinant viral genome by in vitro transposition. Moreover, the extracellular RNase gene from Bacillus amyloliquefaciens, barnase, was expressed under the control of the Cotesia plutellae bracovirus early promoter to negatively select against the non-recombinant background. The bEasyBac bacmid could only replicate in host insect cells when the barnase gene was replaced with the gene of interest by in vitro transposition. When bEasyBac was transposed with pDualBac-EGFP, the resulting recombinant virus, AcEasy-EGFP, showed comparable levels of EGFP expression efficiency to the plaque-purified recombinant virus AcEGFP, which was constructed using the bAcGOZA system. In addition, no non-recombinant backgrounds were detected in unpurified AcEasy-EGFP stocks. Based on these results, a high-throughput system for the generation of multiple recombinant viruses at a time was established.</P>
Choi, Jae Young,Roh, Jong Yul,Li, Ming Shun,Shim, Hee Jin,Kang, Joong Nam,Woo, Soo Dong,Jin, Byung Rae,Je, Yeon Ho Korean Society of Sericultural Science 2004 International Journal of Industrial Entomology Vol.9 No.2
Recently, we have developed an easy, simple and convenient circular DNA cloning system named plasmid capture system (PCS). To investigate usefulness of PCS in cloning of plasmids from Bacillus thuringiensis strains, PCS donors, pPCS-S and pPCS-L were applied to clone plasmids of B. thuringiensis subsp. israelensis by in vitro transposition using 4{TnsABC^*}$ transposase. In result, 3 small plasmids were cloned, and these were consistent with pTX14-1, pTX14-2 and pTX14-3 reported previously from B. thuringiensis subsp. israelensis. Therefore, the PCS can be successfully applied to clone small plasmids from B. thuringiensis strains.
Choi, Woo-Sung,Jang, Myeong Je,Park, Yoo Sei,Lee, Kyu Hwan,Lee, Joo Yul,Seo, Min-Ho,Choi, Sung Mook American Chemical Society 2018 ACS APPLIED MATERIALS & INTERFACES Vol.10 No.45
<P>We prepare three-dimensional honeycomb-like Cu<SUB>0.81</SUB>Co<SUB>2.19</SUB>O<SUB>4</SUB> nanosheet arrays supported by Ni foam via electrochemical codeposition of cobalt and copper hydroxides on Ni foam followed by thermal oxidation. The codeposition with Cu changes the morphology of the cobalt hydroxide deposit to form honeycomb-like nanostructures, significantly decreasing the onset potential for oxygen evolution. The Cu<SUB>0.81</SUB>Co<SUB>2.19</SUB>O<SUB>4</SUB> anode displays an exceptionally low overpotential of 290 mV at a current density of 10 mA cm<SUP>-2</SUP> in 1 M KOH, and an anion-exchange membrane water electrolysis cell employing the above anode achieves a current density of 100 mA cm<SUP>-2</SUP> at 1.68 V in 0.1 M KOH.</P> [FIG OMISSION]</BR>
Analysis of promoter activity of selected Cotesia plutellae bracovirus genes
Choi, Jae Young,Kwon, Soo-Jin,Roh, Jong Yul,Yang, Tae-Jin,Li, Ming Shun,Park, Beom-Seok,Kim, Yonggyun,Woo, Soo-Dong,Jin, Byung Rae,Je, Yeon Ho Microbiology Society 2009 The Journal of general virology Vol.90 No.5
<P>In a previous study, we cloned 27 discrete genome segments of Cotesia plutellae bracovirus (CpBV) and provided the complete nucleotide sequences and annotation. Seven putative coding regions were predicted from one of the largest segments, CpBV-S30. The activity of promoters associated with six predicted ORFs from this segment were investigated using both transient and baculovirus expression assays with enhanced green fluorescent protein as a reporter gene. CpBV promoters showed activity earlier than the polyhedrin promoter and the activity of some of these promoters was superior to that of the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ie-1 promoter in the baculovirus expression assays. The promoter of ORF3004 showed the highest level of activity in insect cells, exhibiting 24 % of the activity obtained with the AcMNPV polyhedrin promoter in Sf9 cells. In Spodoptera exigua larvae, the ORF3006 promoter showed the highest activity, with about 35 % of the activity measured with the polyhedrin promoter. In addition, analysis of the ORF3006 promoter revealed that the region between -382 and -422 from the translation start point was critical for activity of this promoter. These results suggest that the CpBV-S30 promoters characterized here could be useful tools in a variety of biotechnological applications, such as gene expression analyses and insecticide development.</P>
Choi, Jae-Young,Wang, Yong,Kim, Yang-Su,Kang, Joong-Nam,Roh, Jong-Yul,Woo, Soo-Dong,Jin, Byung-Rae,Je, Yeon-Ho 한국응용곤충학회 2008 Journal of Asia-Pacific Entomology Vol. No.
To investigate the usefulness of early-expressed promoters from Cotesia plutellae bracovirus(CpBV) in improvement of insecticidal activities of baculovirus, recombinant AcNPVs, Ac3003ProAalT, Ac3004ProAalT, Ac3005ProAaIT and Ac3006ProAalT expressing scorpion neurotoxin, AaIT, under the control of ORF3003, ORF3004, ORF3005 and ORF3006 promoter, respectively were constructed. Among these recombinant viruses, Ac3006ProAaiT showed highest insecticidal activity against 3rd instar larvae of Spodoptera exigua. These results suggested that early promoters from CpBV could be successfully applied to improve pathogenicity of baculoviruses.