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들민달팽이(Deroveras varians)의 배자발육과 난황단백질
진병래,손흥대,박혜진,조은숙 東亞大學校附設 農業生命科學硏究所 1999 農業生命資援硏究 Vol.8 No.1
들민달팽이의 배자발육은 해부현미경으로 관찰하였다. 산란 후 4일이 경과면서 발육이 급격하게 진전되어, 10일이 지나면서 난 내에서 들민달팽이 유충의 형태를 갖추고, 12일째 부화하였다. 들민달팽이의 배자발육에 따른 난황단백질의 변화는 산란 1일째, 3일째 및 6일째 알의 난황단백질을 전기영동방법으로 분석하였다. 그 결과 배자발육과 함께 급격히 감소되는 난황단백질 관련 밴드를 관찰할 수 있었다. We characterized embryogenesis and egg proteins of Variable field Slug, Deroceras varians. The embryogenesis was observed by light microscopy. The eggs were hatched at the twelfth day after oviposition. The egg proteins of the first day, third day and sixth day after oviposition were analyzed by native-and SDS-PAGE analyses. The results showed that yolk proteins are gradually decreased during embryogenesis.
이대원,박현우,진병래,정영호,박영목,강석권 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.2
미생물 살충제로 사용되고 있는 B. thuringiensis를 모기유충방제에 적용하기 위한 시도는 B. thuringiensis가 수서생태계에서 부유하지 못하고 가라앉으며, 생활환을 이루지 못한다는 문제점으로 인해 적용에 어려움이 있다. 따라서 본 연구는 모기유충에 강한 독성을 갖는 B. thuringiensis subsp. morrisoni PG-14의 cryIVD 유전자를 모기유충의 먹이인 cyanobactrium에 도입하기 위하여 발현벡터 pCYASK 5-1을 제작하고, cyanobacterium Synechocystis PCC6803에 형질전환시켜, 세포내에서 cryIVD 유전자의 발현과 뇌염모기에 대한 독성을 조사하였다. 그 결과 형질전환체 내에서 cryIVD 발현은 immunoblot 분석을 통해 B. thuringiensis subsp. morrisoni PG-14에서 발현된 단백질과 같은 분자량으로 발현되는 것을 확인하였으며, 모기유충에 대한 형질전환체의 독성은 1.40×10^6 cells/ml 농도에서 약 80%의 치사율을 나타내었다. 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 Synechocystis PCC6803 reported as a natural mosquito larval food source and the transformants were selected with kanamycin. Expression of cryIVD gene in transformant was characterized by SDS-polyacrylamide gel electrophoresis (PAGE) and immunoblot analysis. The mosquitocidal activity of a transformant was determined 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.
학교체육 활성화 방안에 대한 연구(Ⅰ) : 강원대학교 교양체육 운영방법의 개선을 중심으로
이광재,문병용,노성규,박기동,홍관이,한상준,유옥재,엄기진,정청자,오수일,김윤래,박장평,부기원 江原大學校附設 體育科學硏究所 1986 江原大學校附設體育科學硏究所論文集 Vol.- No.11
Try to find a scheme for activation to College Physical Education, this Report informs the various kinds of Survey and process those were projected for improvement of management methods on Cultural Physical Education in Kang Weon National University. From experimentally carried out "Sports events Choosing System" not as usual be inforced "Total practical Skill System", Following Positive effects and sujestions were Presented; 1. "Sports events choosing system" gave birth to more students' interesting and more voluntary participation to the sports events they chose, and sports skill and instruction level was elevated due to charge of expert according to each sports events. This system would be managed continuously hereafter for it related to life sports. 2. This System needs to closed cooperations between College and Community, for the facilities of community are utilized by college. 3. To complete the plan for activation of college Physical Education, additionally, the successional study ; namely "autonomous extracurricular sports activity" should be excuted hereafter.
새로운 Baculoviurs 전이벡터를 이용한 Escherichia coli β-galactosidase 유전자의 발현
우수동,김우진,김혜성,진병래,강석권 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1
국내에서 분리된 BmNPV를 이용하여 제작된 새로운 전이벡터 pBmKSK1에 외래 유전자로서 E. coli lacZ 유전자를 클로닝하고 재조합 바이러스를 제작하였다. 재조합 바이러스에 대하여 Southern blotting 분석을 실시하여 lacZ 유전자의 존재를 확인하였으며, 재조합 바이러스가 접종된 세포의 SDS-PAGE 및 Western blotting 분석을 통하여 β-galactosidase의 발현을 확인하였다. 재조합 바이러스의 발현효율을 누에 세포주 및 유충에서 비교 조사한 결과, 유충에서 더욱 높은 발현량을 보임을 확인하였다. 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 coninfected with transfer vector pBmKSK1-LacZ and vild type BmNPV genome, and analysed by Southern blotting. The expression of β-galactosidase was characterized by SDS-PAGE, Western blotting and β-galactosidase activity assay. The results showed that the level of expression in silkworn larvae was higher than that of BmN-4 cells.
cDNA Sequence and mRNA Expression of a Novel Peroxiredoxin from the Firefly, pyrocoelia rufa
Jin, Byung-Rae,Lee, Kwang-Sik,Kim, Seong-Ryul,Sohn, Hung-Dae Korean Society of Sericultural Science 2002 International Journal of Industrial Entomology Vol.4 No.2
We describe here the cDNA sequence and mRNA expression of a novel family of the antioxidant protein, peroxiredoxin, from the firefly, Pyracoetia ruin. The 555 bp cDNA sequence codes for a 185 amino acid protein with a calculated molecular mass of approximately 21 kDa. The deduced protein of P. rufa peroxiredoxin gene contains two conserved cysteine residues. Alignment of the deduced protein of P. rufa peroxiredoxin gene showed 71.1% protein sequenceidentity to known insect Drosophila melanogaster peroxiredoxin. Northern blot analysis revealed that the P. rufa peroxiredoxin is specifically expressed in the fat body of P. rufa larvae.
Jin, Byung Rae,Yoon, Hyung Joo,Kang, Seok Kwon,Choudary, Prabhakara V. 東亞大學校附設遺傳工學硏究所 1998 遺傳工學硏究 Vol.- No.5
Genomic DNA of recombinant AcNPV expressing B-galactosidase was cotransfected with p143helicase gene of BmNPV into Sf21 celis. Ac-Bm hybrid viruses capable of replicating in both Bm5 and Sf21 cells were isolated. Ac-Bm hybrid viruses expressing B-galactosidase either at the highest (Ac-Bm hybrid virus-HE) or lowest (Ac-Bm hybrid virus-LE) level were chosen for the characterization of B-galactosidase expression in Bm5 and Sf21 cells. Expression level of B-galactosidase and replication of Ac-Bm hybrid virus-He in Sf21 cells were nearly identical to those of recombinant AcNPV. Furthermore, replication of Ac-Bm hybrid virus-HE in Bm5 cells was similar to that of wild-type BmNPV, and Ac-Bm hybrid virus-HE clearly expressed B-galactosidase in Bm5 cells. However, expression of B-galactosidase by Ac-Bm hybrid virus-HE in Bm5 cells was significantly lower than that expressed in Sf21 cells. The titer or Ac-Bm hybrid virus-HE determined by plaque assays in Bm5 cells was similar to that determined in Sf21 cells, but the plaque size formed by Ac-Bm hybrid virus-HE in Bm5 cells was apparently smaller than that formed in Sf21 cells. In addition, expression levels and virus titers of Ac-Bm hybrid virus-LE in Sf21 and Bm5 were significantly lower than those of Ac-Bm hybrid virus-HE. Therefore, DNA sequences were determined for the region of the p143 gene controlling the host range in Ac-Bm hybrid viruses. The results showed that the deduced amino acid sequences of Ac-Bm hybrid virus-LE were different at position 461, 470, 514, and 528 from those of BmNPV. In conclusion, our results clearly demonstrated that Ac-Bm hybrid virus-HE has an additional advantage of expanded host range for producing recombinant proteins.
Jin, Byung-Rae,Yun, Eun-Young,Kang, Seok-Woo,Yoon, Hyung-Joo,Kim, Keun-Young,Kim, Ho-Rak,Je, Yeon-Ho,Kang, Seok-Kwon Korean Society of Sericultural Science 2000 International Journal of Industrial Entomology Vol.1 No.2
We have constructed a recombinant baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV), containing green fluorescent protein (GFP) gene from the jellyfish, Aequorea victoria, and transferred it into the domestic silkworm Bombyx mori larvae for the production of visible transgenic silkworm of living organism. When one day-old fifth instar female larvae were injected with the recombinant AcNPV of 1x10$^{5}$ plaque forming units, the bright glow of GFP was detected in the recombinant AcNPV-infected larvae and in the newly hatched larvae of the next generation. Our findings demonstrate that the viral replication was detected in the silkworm treated with the recombinant ACNPV and the gfp gene was expressed under the transcriptional control of the polyhedrin gene promoter, Furthermore, the gfp gene was transmitted to the next generation, suggesting that this system can be applied for the development of transgenic silkworms.