• Journal of Microbiology and Biotechnology
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• v.25 no.3
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• pp.386-392
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• 2015

A novel recombinant bacmid, bEasyBm, that enables the easy and fast generation of pure recombinant baculovirus without any purification step was constructed. In bEasyBm, 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 nonrecombinant background. The bEasyBm bacmid could only replicate in host insect cells when the barnase gene was replaced with the gene of interest by in vitro transposition. When bEasyBm was transposed with pDualBac-EGFP, the resulting recombinant virus, EasyBm-EGFP, showed high levels of EGFP expression efficiency compared with that of non-purified recombinant virus BmGOZA-EGFP, which was constructed using the bBmGOZA system. In addition, nonrecombinant backgrounds were not detected in unpurified EasyBm-EGFP stocks. Based on these results, a high-throughput system for the generation of multiple recombinant viruses at a time was established.

• Journal of Sericultural and Entomological Science
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• v.41 no.2
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• pp.102-107
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• 1999

To analysis a promoter strength of Atographa californica nucler polyhedrosis virus (AcNPV) IE1 gene, an immediate viral gene, ${\beta}$-glactosidase gene as a reporter gene was introduced under the control of the IE1 promoter. The restriction fragment containing IE1 promoter and ${\beta}$-galctosidase gene from pAcIE1-gal were inserter into pBacPAK9 to yield transfer vector pAcNPV-IE1-gal. The pAcNPV-IE1-gal was cotransfected with AcNPV genomic DNA BacPAK6 into Sf9 cells to produce recombinant baculovirus AcNPV-IE1-gal. In addition, recombinant bacvulovirus AcNPV-gal, which express ${\beta}$-galac-tosidase under the control of the polyhedrin promoter, was constrer, was constructed to compared with AcNPV-IE1-gal. The recombinant viruses were respectively infected into Sf9 cells and characterized by the virus titer and expression of ${\beta}$-galactoxidase in Sf9 cells. The promoter strength of IE1 and polyhedrin promoters was determined by the amount of ${\beta}$-galactosidase secreted into medium by viral infection. The titer of AcNPV-IE1-Gal determined by plaque assays in Sf9 cells was similar to that of AcNPV-gal. However, expression level of ${\beta}$-galactosidase by AcNPV-IE1-gal was significantly lower than that by AcNPV-gal. In conclusion, promoter strength of IE1 was approximately 25-fold lower than that of polyhedrin.

• The Plant Pathology Journal
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• v.27 no.1
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• pp.37-43
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• 2011

Rice stripe virus (RSV) is one of serious epidemic pathogens for rice species grown in many Asian countries. Therefore, it is necessary to produce a diagnostic detection kit applicable in fields for RSV detection. In this study, RSV proteins that were derived from recombinant proteins and synthetic polypeptides as antigens were generated and were raised in rabbits for antiserum production. Among seven proteins in RSV, genes that code for NCP and NS3 proteins were cloned and subcloned into vector carrying His-tag protein and were expressed in E. coli. Of two recombinant proteins, only anti-NCP displayed stable hybridization signals in western blot analysis. Alternately, synthetic RSV polypeptides for CP, NCP, NS3 and NSvc4 we also generated and only antibodies against CP and NCP were very effective to detect RSV in both RSV infected rice and weed plants. However, antibodies against NS3 and NSvc4 showed weak specific bands as well as strong non-specific background due to the difference of viral proteins produced in the infected leaves. In summary, the antibodies generated against RSV proteins produced in this study will be useful for various assays such as for RSV diagnostic detection, immunoprecipitation, protein purification, and western blot analysis.

• The Plant Pathology Journal
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• v.36 no.3
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• pp.280-288
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• 2020

RNA interference (RNAi) has attracted attention as a promising approach to control plant viruses in their insect vectors. In the present study, to suppress replication of the rice stripe virus (RSV) in its vector, Laodelphax striatellus, using RNAi, dsRNAs against L. striatellus genes that are strongly upregulated upon RSV infection were delivered through a rice leaf-mediated method. RNAi-based silencing of peroxiredoxin, cathepsin B, and cytochrome P450 resulted in significant down regulation of the NS3 gene of RSV, achieving a transcriptional reduction greater than 73.6% at a concentration of 100 ng/μl and, possibly compromising viral replication. L. striatellus genes might play crucial roles in the transmission of RSV; transcriptional silencing of these genes could suppress viral replication in L. striatellus. These results suggest effective RNAi-based approaches for controlling RSV and provide insight into RSV-L. striatellus interactions.

• Journal of Life Science
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• v.12 no.1
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• pp.49-54
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• 2002

Development of a rapid method possessing the requisite sensitivity and specificity for virus monitoring is necessary for protection of the shellfish-consuming public. Oysters tissue usually contains virus particles in relatively small concentrations along with various other substances that can interfere with detection steps. Therefore, the critical point concerning the detection of viruses in shellfish tissues resides in the processing of samples. The current study demonstrated the possibility of purifying small amounts of virus particles at the interface of a 10/50% sucrose gradient after a single round of sucrose gradient ultracentrifugation. We could detect HAV and poliovirus simultaneously from oyster tissues by using two different sets of primer. Furthermore, the method showed a high level of virus recovery rate (>95%) as determined by plaque assays of the final samples. Taken the advantages of the simple and sensitive methods, it was possible to detect 2 pfu of HAV in 5 g of oyster digestive tissues within 24h.

• Journal of the Korea Institute of Information Security & Cryptology
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• v.15 no.6
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• pp.37-47
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• 2005

Recently, the malicious e-mail worm-viruses have been widely spreaded over the Internet. If the recipient opens the e-mail attachment or an e-mail itself that contains the worm-virus, the worm-virus can be activated and then cause a tremendous damage to the system by propagating itself to everyone on the mailing list in the user's e-mail package. In this paper, we have designed and implemented two methods blocking e-mail worm-viruses. In the fist method, each e-mail is transmitted only by sender activity such as the click of button on a mail client application. In the second one, we insert the two modules into the sender side, where the one module transforms a recipient's address depending on a predefined rule only in time of pushing button and the other converts the address reversely with the former module whenever an e-mail is sent. The lader method also supports a polymorphism model in order to cope with the new types of e-mail worm-virus attacks. The two methods are designed not to work for the e-mail viruses. There is no additional fraction on the receiver's side of the e-mail system. Experimental results show that the proposed methods can screen the e-mail worm-viruses efficiently with a low overhead.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.2
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• pp.127-131
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• 2004

A low-cost, simple strip reader system using a linear movement mechanism of CD-ROM deck has been developed to characterize a lateral flow membrane-based immunochromatographic assay. The test strip reader was assembled by a CD-ROM deck and home-made optical head especially designed for immunoassays. The optical head for detecting reflected light from the test strip surface consists of green light-emitting diode, large area silicon photodiode, and anodized aluminum mounting block providing a slit structure for cutting light from the LED. The stepping motor of the deck was operated in the full step mode, whose distance of each reading point is about 0.15mm. The performance of the strip reader was tested by analysis of HBV(hepatitis B virus) antigen test kit. This strip reader can be useful for inexpensive, disposable, and membrane-based assays that provide visual evidence of the presence of an analyte in a liquid sample.

• Proceedings of the Korean Information Science Society Conference
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• 2005.11b
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• pp.262-264
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• 2005

인유두종 바이러스(Human Papillomavirus: HPV)는 사마귀로부터 생식기 및 배설기의 침윤성 암에 이르기까지 여러 질병과 연관되어 있음이 알려져 있다. 현재 200종 이상이 알려져 있고, 이 중 85개는 전체 유전자가 밝혀져 있다. HPV 감염 시 만들어지는 단백질 중 E6. E7 단백질은 암 억제 유전자(p53, pRb)에 결합하여 세포의 암 억제 기능을 저하시키고 이로 인해 암을 발생시킨다. 본 논문은 암 발생과 밀접한 관련이 있는 HPV의 E6 단백질 서열과 HPV 유형(HPV Type)을 가지고, PLSA (Probabilistic Latent Semantic Analysis) 방법을 이용하여 HPV를 클러스터링(clustering) 해 보았다. 실험 결과, 특정 클러스터는 질병과 밀접하게 연관되어 있으며, 이와 관련된 주요 서열 분석이 가능함을 보여주고 있다.

• Pediatric Infection and Vaccine
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• v.4 no.1
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• pp.116-125
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• 1997

Purpose : Studies on the duration of immune response against Japanese encephalitis virus from recipients with JE vaccine (Nakayama-NIH strain) in Korea. Methods : To determinate the immune response and the duration of antibody against JE vaccine, 213 students were examined since 1994 using hemmaglutination inhibition test and plaque reduction neutralization test (PRNT). Results : 24 months after the first vaccination, haemmaglutination inhibition and neutralizing antibody maintained from the recipients 63.4% (>1:20) and 100% (>1:20), respectively. In April 1996, one dose booster to the same recipients those who were vaccinated in 1994, the GMT antibody for HI and PRNT titer were both increased from 1:11.6 to 1:13.2 and 1:275.7 to 1:348.1, respectively, after 6 months booster (after 30 months from the initial vaccination). This results showed that the antibody from the active immunity could be maintained more than 12 months after the initial vaccination. On the basis of these results, inactivated killed JE vaccine (Nakayama-NIH strain) using for preventing against JE purpose seems to produce antibody enough to protect against JE at present. Conclusions : Along with the results of this study demonstrating duration of antibody, the active immunization could be maintained as long as by initial vaccination of 2 doses, a single dose of booster vaccination made during a period of 1 month to 12 months and the successive booster vaccination by 2 or 3 year intervals. However, the immunization schedule should be concerned with both epidemiology of disease and the immune response of vaccinated individuals.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.2
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• pp.149-153
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• 2000

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.