• 식물병연구
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• 제10권1호
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• pp.30-33
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• 2004

현재까지 벼 줄무의잎마름병(Rice stripe virus, RSV)은 남부지방에 국한되어 발생되어 왔다. 그러나 최근에는 RSV의 발생이 충청도, 경기도를 포함한 중부지방까지 확산되는 경향을 나타내고 있다. 이병의 병징은 육안으로는 생리적인 장해 현상과 구분하기가 힘들다 본 실험에서는 이병주 및 애멸구(Laodelphax striatellus)로 부터 viral RNA를 추출한 후 RNA복제효소 및 외피단백질유전자에 특이적인 primer를 제작하여 RT-PCR법에 의해 RSV를 검정하였다. 그결과 이병식물체 및 보독 애멸구로부터 RNA복제효소 유전자에 특이적인 band(1,023 bp) 및 외피단백질유전자에 특이적인 band(969 bp)가 관찰되었다.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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• pp.82-83
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• 2003

In previous experiment, we reported when the heterologous protein is expressed by using baculovirus expression vector system (BEVS), although the amount of intracellular protein is abundant, the amount of extracellular Protein is poor. As the link in the chain of the research, we investigated the secretory pathway, important in case of the secretory protein, of the protein expressed in insect cells using BEVS. (omitted)

• 한국산림과학회지
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• 제101권4호
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• pp.635-639
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• 2012

뽕나무 오갈병 파이토플라스마 매개곤충을 알아보기 위하여 뽕나무 재배지에서 채집된 곤충 중 흡즙성 곤충 3 종(Hishimonas sellatus, Bothrogonia japonica and Balclutha punctata)를 선발하였다. 이들 흡즙성 곤충을 뽕나무 이병주에서 흡즙시킨 다음 건전 뽕나무와 일일초로 전반시킨 결과, 마름무늬매미충(H. sellatus)과 B. punctata이 파이토플라스마를 전반하였다. 뽕나무에서는 전형적인 오갈 증상을 보였지만 일일초에서는 가지가 가늘게 늘어지며, 잎은 총생하고 작아지는 빗자루증상이 나타났다. 매개충과 매개전염시킨 뽕나무와 일일초에 대하여 R16F2n/R2 프라이머로 PCR 검정 결과 끝검은말매미충을 제외한 모든 시료에서 파이토플라스마가 검출되었다.

• Reproductive and Developmental Biology
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• 제33권2호
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• pp.71-76
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• 2009

Salivary lipocalin (SAL1) is a member of the lipocalin protein family that has a property to associate with many lipophilic molecules. The importance of SAL1 during pregnancy in pigs has been suggested by our previous study which has shown that SAL1 is expressed in the uterine endometrium in a cell type- and implantation stage-specific manner and secreted into the uterine lumen. However, function of SAL1 in the uterus during pregnancy in pigs is not known. To understand SAL1 function in the uterus during pregnancy, we generated recombinant porcine SAL1 protein in an insect cell line. Porcine SAL1 cDNA was cloned into a baculovirus expression vector using RT-PCR and total RNA from uterine endometrium on day 12 of pregnancy, and the expression vector was used to generate recombinant Bacmid containing the SAL1 gene. The recombinant Bacmid was then transfected Sf9 cell to produce recombinant baculovirus. By infecting Sf9 cell with recombinant baculovirus, we established a SAL1-expressing insect cell expression system. Immunoblot analysis confirmed SAL1 expression in the infected cells. Recombinant SAL1 produced by the Sf9 cell line will be useful for understanding physiological function of SAL1 during pregnancy in pigs.

• BMB Reports
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• 제33권6호
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• pp.483-492
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• 2000

The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

• 대한바이러스학회지
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• 제29권2호
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• pp.137-144
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• 1999

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.

• 대한바이러스학회지
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• 제28권3호
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• pp.225-232
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• 1998

We have expressed GFP in Sf9 and Bm5 cells or Bombyx mori larvae by using Ac-Bm hybrid virus capable of replicating in both Bm5 and Sf9 cells. Genomic DNA of Ac-Bm hybrid virus expressing ${\beta}$-galactosidase was cotransfected with baculovirus transfer vector containing GFP gene, pBacPAK-GFP in Sf9 cells. The Ac-Bm hybrid virus harboring GFP was named as Ac-Bm hybrid virus-GFP. The Ac-Bm hybrid virus-GFP-infected insect cells were easily selected by detecting the emission of GFP from each well of cell culture dish on the UV illuminator. GFP produced by Ac-Bm hybrid virus-GFP in Sf9 and Bm5 cells or B. mori larvae was confirmed by SDS-PAGE and Western blot analysis using GFP antibody. In addition, B. mori larvae infected with Ac-Bm hybrid virus-GFP was apparently appeared fluorescence from the whole body at S days postinoculation. The fluorescence of GFP from the hemolymph and fat body of B. mori larvae infected with Ac-Bm hybrid virus-GFP was also observed by fluorescence microscope. In conclusion, our results demonstrated that in baculovirus expression vector system, use of Ac-Bm hybrid virus have an additional advantage of expanded host range for producing recombinant proteins.

• 한국식물병리학회지
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• 제1권3호
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• pp.190-194
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• 1985

벼검은줄오갈병 매개충인 애멸구의 기주식물 선호성은 실내유균접종과 포장상태에서 공히 옥수수가 벼보다 낮으나 발병율은 아주 높았다. 바이러스가 매개충의 생태에 미치는 영향은 보독충이 무독충보다 성충의 생존일수, 부화약충수 및 성충율에서 심한 장애를 나타냈다. 벼에서는 감염초기부터 후기까지 건물중이 이병주가 건전주보다 낮았다. 그러나 옥수수에서는 감염초기에 이병주가 건전주보다 건물중이 높았다. 한천내확산법 및 미량심강법에 의한 감염바이러스의 정량결과, 항혈청과의 반응정도는 옥수수잎, 벼줄기, 벼잎, 옥수수줄기에서 부분순화한 항원바이러스의 순으로 나타났다. 이병기주식물의 phloem gall 형성에서 저온처리의 효과가 인정되었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.142-146
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• 2003

Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

• BMB Reports
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• 제34권4호
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• pp.371-378
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• 2001

The Glycoprotein D (gD) gene of the HSV-1 strain F was cloned, sequenced, recombinated into the HcNPV (Hyphantria cunea nuclear polyhedrosis virus) expression vector and expressed in insect cells. The gD gene was located in the 6.43 kb BamHI fragment of the strainF. The open reading frame (ORF) of the gD gene was 1,185 by and codes 394 amino acid residues. Recombinant baculoviruses, GD-HcNPVs, expressing the gD protein were constructed. Spodoptera frugiperda cells, infected with the recombinant virus, synthesized a matured gX-gD fusion protein with an approximate molecular weight of 54 kDa and secreted the gD proteins into the culture media by an immunoprecipitation assay The fusion gD protein was localized on the membrane of the insect cells, seen by using an immunofluorescence assay The deduced amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. These results indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins.