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이대원,박현우,진병래,정영호,박영목,강석권,Lee, Dae-Weon,Park, Hyun-Woo,Jin, Byung-Rae,Chung, Young-Ho,Park, Young-Mok,Kang, Seok-Kwon 한국미생물 · 생명공학회 1996 한국미생물·생명공학회지 Vol.24 No.2
Bacillus thuringiensis subsp. morrisoni PG-14 is a gram-positive soil bacterium producing mosquitocidal parasporal inclusions composed of several crystal proteins. Among these crystal protein genes, cryIVD gene is one of major component which has 72 kDa in size. However, these parasporal inclusions sink quickly from the surface of water where mosquito larval feeding occurred. To develope mosquitocidal cyanobacterium, therefore, we constructed the expression vector, pCYASK 5-1 harboring cryIVD gene. The expression vector, pCYASK5-1 was transformed into the cyanobacterium Syne- chocystis PCC6803 reported as a natural mosquito larval food source and the transformants were selected with kanamycin. Expression of IVD gene in transformant was characterized by SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblot analysis. The mosquitocidal activity of a transformant was determined with Culex tritaeniorhynchus. The results showed that, the transformed cyanobacterium is toxic to mosquito larvae and will be expected as a potential agent that is used for mosquito control.
A stydy on The Biological Control of Sciarid Fly(Lycoriella sp.) Using Bacillus thuringiensis
최광호,박현철,박현우,진병래,강석권,손흥대,Choi, Kwang-Ho,Park, Hyean-Cheal,Park, Hyun-Woo,Jin, Byung-Rae,Kang, Seok-Kwon,Sohn, Hung-Dae 한국생명과학회 1996 생명과학회지 Vol.6 No.4
Thirteen subspecies of Bacillus thuringiensis including B. t. israelensis, B. t. morrisoni PG-14 and B.t. darmstadoemsos known to be toxic to dipteran insects were treated on the mushroom (Flammulina velutipes) compost to estimate the biological control effect of a sciarid fly, Lycoriella sp. According to the results, it was found that there were no significant effects of the tested strains of B, thuringiensis on the control of Lycoriella sp. For further confirmation, larval gut juice of Lycoriella sp. and trypsin were respectively treated into the parasporal crystal proteins of three subspecies of B. t. israelensis, B. t. morrisoni PG-14, and B. t. darmstadiensis. The proteins were separated by SDS-PAGE. According to the results, the major parasporal crystal proteins were respectively produced by B. t. morrisoni as the amount of 52 kd, B. y. israelensis as 37kd and B. t. darmstadiensis as 39kd, but the activity of these proteins could not be unfortunately confirmed in this study.
제연호(Yeon Ho Je),진병래(Byung Rae Jin),노종열(Jong Yul Roh),장진희(Jin Hee Jang),강석권(Seok Gwon Kang) 대한바이러스학회 1999 Journal of Bacteriology and Virology Vol.29 No.3
We have now constructed a novel recombinant baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) producing polyhedra with Bacillus thuringiensis (Bt) CryIAc crystal protein. The recombinant polyhedra produced by the recombinant baculovirus, Btrus, in insect cells was characterized. The recombinant baculovirus has two independent transcription units in opposite orientations with two promoters, p10 or polyhedrin gene promoter each initiating transcription of either native polyhedrin or fusion protein with polyhedrin and Bt CrylAc crystal protein. Surprisingly, this recombinant baculovirus stably produced recombinant polyhedra which were nearly similar to those of wild-type AcNPV. The immunogold staining experiment showed that the recombinant polyhedra were assembled with polyhedrin and Bt Cry1Ac crystal protein, and contained virus particles. Insecticidal toxicity of recombinant polyhedra of Btrus to the faU webworm, Hyphantria cunea, was strikingly improved in comparison with the wild-type AcNPV.
새로운 Baculovirus 전이벡터를 이용한 Escherichia coli ${\beta}$-galactosidase 유전자의 발현
우수동,김우진,김혜성,진병래,강석권,Woo, Soo-Dong,Kim, Woo-Jin,Kim, Hye-Seong,Jin, Byung-Rae,Kang, Seok-Kwon 한국미생물 · 생명공학회 1996 한국미생물·생명공학회지 Vol.24 No.1
To investigate the expression efficiency of new transfer vector of Bombyx mori nuclear polyhedrosis virus (BmNPV), Escherichia coli lacZ gene was inserted into new transfer vector pBmKSK1, under the control of polyhedrin promoter and expressed in BmN-4 cells and larvae of silkworm, Bombyx mori. The recombinant virus containing lacZ gene was isolated from BmN-4 cells coinfected with transfer vectro pBmKSK1-LacZ and wild type BmNPV genome, and analysed by Southern blotting. The expression of ${\beta}$-galactosidase was characterized by SDS-PAGE, Western blotting and ${\beta}$-galactosidase activity assay. The results showed that the level of expression in silkworm larvae was higher than that of BmN-4 cells.
Autographa californica 핵다각체병 바이러스의 IE1 유전자에 의해 형질전환된 곤충세포주의 특성
조은숙,박혜진,이광식,강석우,윤은영,김근영,손흥대,진병래,Cho, Eun-Sook,Park, Hae-Jin,Lee, Kwang-Sik,Kang, Seok-Woo,Yun, Eun-Young,Kim, Keun-Young,Sohn, Hung-Dae,Jin, Byung-Rae 대한미생물학회 1999 Journal of Bacteriology and Virology Vol.28 No.1
Transformation efficiency, virus multiplication and foreign gene expression were characterized in the insect cells transformed with Autographa californica nuclear polyhedrosis virus (AcNPV) immediate early 1 gene (IE1). Transformation efficiency of insect cells by AcNPV IE1 gene vector horboring foreign gene was approximately 8-fold higher in the Sf9 cells transformed previously with AcNPV IE1 gene than in the normal Sf9 cells. Virus multiplication and foreign gene expression of recombinant baculovirus in the Sf9 cells transformed with AcNPV IE1 gene were similar to those of the normal Sf9 cells. These results suggest that transformed cells displaying foreign gene product by using AcNPV IE1 gene promoter will be useful for the diverse applications of insect cells.
Autographa californica 핵다각체병 바이러스의 IE1 유전자에 의해 형질전환된 곤충세포주의 특성
조은숙(Eun Sook Jo),박혜진(Hye Jin Park),이광식(Gwang Sik Lee),강석우(Suk Woo Kang),윤은영(Eun Young Yoon),김근영(Geun Young Kim),손홍대(Hong Dae Son),진병래(Byung Rae Jin) 대한바이러스학회 1999 Journal of Bacteriology and Virology Vol.29 No.2
Transformation efficiency, virus multiplication and foreign gene expression were characterized in the insect cells transformed with Autographa californica nuclear polyhedrosis virus (AcNPV) immediate early 1 gene (lE1). Transformation efficiency of insect cells by AcNPV IE1 gene vector horboring foreign gene was approximately 8-fold higher in the Sf9 cells transformed previously with AcNPV IE1 gene than in the normal Sf9 cells. Virus multiplication and foreign gene expression of recombinant baculovirus in the Sf9 cells transformed with AcNPV IE1 gene were similar to those of the normal Sf9 cells. These results suggest that transformed cells displaying foreign gene product by using AcNPV IE1 gene promoter will be useful for the diverse applications of insect cells.