• 한국식물병리학회지
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• 제14권5호
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• pp.513-518
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• 1998

Beet curly top virus (BCTV) mutant has been constructed in vitro that contain G-to-T transversions at nucleotide 2727 within overlapping open reading frames (ORFs) L1 and L4. The mutations introduce termination codon in ORF L4 without affecting the amino acid encoded by ORF L1. When agroinoculated into Arabidopsis thaliana the mutant caused mild stunting and stem curling, but not the callus induction and hyperlasia on infected tissues of Sei-O ecotype. However, this mutant was not infectious on Col-O. Levels of single stranded DNA forms were similar in mutant and wild type BCTV infections. The DNA quantitation data showed that the DNA of BCTV-L4 mutant virus was accumulated in shoot tips, infection origin and roots with similar levels to those of wild type virus infected. Three tissues of asymptomatic ecotype Col-O also had as much as virus DNA from wild type virus infections. In both ecotypes infected with BCTV-Logan and BCTV-L4 mutant, root tissues contained more virus DNA than any other tissues by the Southern hybridization data. The results suggest that ORF L4 encodes a functional protein that is a major determinant of pathogenesis that might affect the hyperplastic response of the host to BCTV infection.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1979년도 추계학술대회 심포지움
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• pp.248.1-248
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• 1979

Sixteen temperature-sensitive mutants of Autographa californica nuclear polyhedrosis virus were isolated. Several interesting phenotypes were observed. A large proportion of the mutants were un-able to form polyhedral occlusion bodies at the nonpermissive temperature (32.5C). At 32.5C, one mutant formed plaques in which the cells lacked polyhedra. Another mutant type was defective in the production of progeny extracellular nonoccluded virus and produced a plaque consisting of only a single cell containing polyhedra at 32.5C. One mutant was defective in plaque formation, progeny nonocluded virus formation, and polyhedra formation at 32.5C. Several mutants produced nonocluded virus but failed to produce plaques or polyhedra at 32.5C. Other phenotypes were also distinguished. Complementation analyses, performed by either measuring the increase in extracellular non-ocludedvirus formation or by oberving polyhedra formation in mixed infections at 32.5C, indicated the presence of 15 complementation groups. A high frequency of recombination was observed. Four of the mutants were found to be host dependent in their temperature sensitivity for polyhedra formation.

• Biomolecules & Therapeutics
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• 제21권2호
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• pp.97-106
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• 2013

Chronic hepatitis C virus (HCV) infection is responsible for the development of liver cirrhosis and hepatocellular carcinoma. HCV core protein plays not only a structural role in the virion morphogenesis by encapsidating a virus RNA genome but also a non-structural role in HCV-induced pathogenesis by blocking innate immunity. Especially, it has been shown to regulate JAK-STAT signaling pathway through its direct interaction with Janus kinase (JAK) via its proline-rich JAK-binding motif ($^{79}{\underline{P}}GY{\underline{P}}WP^{84}$). However, little is known about the physiological significance of this HCV core-JAK association in the context of the virus life cycle. In order to gain an insight, a mutant HCV genome (J6/JFH1-79A82A) was constructed to express the mutant core with a defective JAK-binding motif ($^{79}{\underline{A}}GY{\underline{A}}WP^{84}$) using an HCV genotype 2a infectious clone (J6/JFH1). When this mutant HCV genome was introduced into hepatocarcinoma cells, it was found to be severely impaired in its ability to produce infectious viruses in spite of its robust RNA genome replication. Taken together, all these results suggest an essential requirement of HCV core-JAK protein interaction for efficient production of infectious viruses and the potential of using core-JAK blockers as a new anti-HCV therapy.

• Asian-Australasian Journal of Animal Sciences
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• 제9권6호
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• pp.679-684
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• 1996

This study evaluates the susceptibility of scaleless mutant chickens to very virulent Marek's disease virus (vvMDV) inoculation. One day old chickens were inoculated subcutaneously with Taiwanese isolates of LTB-1 and LTS-1 strains, and standard strain of Md/5. Compared with the non-inoculated group the vvMDV-inoculated chickens showed decreased body weights and atrophy of lymphoid organs before 35 days old. These results indicate that scaleless chickens show the same susceptibility as the wild type chickens to vvMDV infection. Furthermore, the protective effect of herpesvirus of turkey (HVT) vaccination at 1 day old against vvMDV challenge was evaluated. Scaleless mutant chickens of treated groups showed 20-30% early death, and 85.7-100% and 12.5-14.2% had lymphomatous lesions in visceral organs and peripheral nerves, respectively. No significant lesions were observed in non-challenged chickens of the control group. The HVT vaccination did not provide an effective protection against vvMDV infection. It is concluded that scaleless mutant chickens are susceptible to vvMDV infection.

• 미생물학회지
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• 제46권2호
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• pp.140-147
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• 2010

일본뇌염바이러스(Japanese encephalitis virus)는 모기 매개성 플라비바이러스에 속하며, 주로 동남아시아 지역에서 유행성 바이러스성 뇌염을 일으킨다. 일본뇌염바이러스는 외피를 가진 작은 바이러스로서, 양성가닥 RNA 게놈을 가지고 있다. 감염성을 띤 바이러스 입자는 capsid (C), membrane (M; prM 전구체로부터 생성), 및 envelope (E)과 같은 3개의 구조단백질로 이루어져 있다. 본 연구에서는 일본뇌염바이러스 생산과 정에 M 단백질의 N-말단부위에 위치한 극성 아미노산 잔기의 역할을 분석하였다. 일본뇌염바이러스의 infectious cDNA를 활용하여, M 단백질의 $E^9$와 $K^{15}K^{16}E^{17}$ 잔기를 알라닌으로 치환시킨 2개의 mutant cDNA (Mm1과 Mm2)를 각각 제작하였다. 각각의 cDNA로부터 합성된 mutant RNA를 세포에 트랜스펙션시킴으로써, 비록 세포 내에 축적된 3개의 구조단백질양은 변화가 없으나, 이들 세포로부터 생산된 바이러스의 양은 Mm2 RNA의 경우 ~1,000배 감소됨을 관찰하였다. 흥미롭게도, Mm2 RNA로부터 발현된 mutant M 단백질은 wild-type M 단백질을 인지하는 항혈청에는 반응하지 않았으나, mutant M 단백질을 항원으로 제작된 항혈청에는 반응하는 것을 알 수 있었다. 본 실험결과는 일본뇌염바이러스 M 단백질을 구성하는 3개의 극성 아미노산 잔기($K^{15}K^{16}E^{17}$)가 바이러스의 생산과정에 관여한다는 것을 암시한다. 앞으로, wild-type 또는 mutant M 단백질(Mm2)을 인식하는 2개의 항혈청은 이 단백질의 기능연구에 유용한 재료로 사용될 것으로 기대된다.

• BMB Reports
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• 제28권4호
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• pp.331-335
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• 1995

Simian virus 40 (SV40) small-t antigen (small-t) has been known to regulate the activity of a cellular enzyme, protein phosphatase 2A (PP2A), composed of A. B, and C subunits, via binding to the A subunit In the study presented here, the stoichiometry of the binding of small-t to PP2A was determined to be 1: 1. It was also shown that small-t binds to the AC form of PP2A with a higher apparent affinity than it binds to the free A subunit. We also characterized the interaction of PP2A with wild-type and various mutant small-ts. A single-point mutant (Val134Met) and a double-point mutant (Trp147Gly;Leu152 Pro) of small-t exhibited 3-fold and 5-fold lower potencies in inhibiting PP2A activity. respectively. This suggests that the region around amino acids between 134 and 152 of small-t might be important in regulating the enzyme activity of PP2A.

• 한국연초학회지
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• 제20권1호
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• pp.57-65
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• 1998

This study was conducted to develop a resistant tobarro against Potato virus Y (PVY) by transformation of the plants with genetically engineered viral genes. The complimentary DNAs (cDNAS) of potato virus Y-necrosis strain (PVY-Vn) replicase mutant genes (3'-deleted, 5'-deleted and ADD-mutant Nlbs) were synthesized through RT-PCR by using purified PVY-VN RNA and synthesized primers, and cloned in the sense orientation into a plant expression vector (pMBPI), The cDNAS of the genes were transferred into Agrobacterium tumefaciens LBA 4404, and then transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Regenerated plants were tested for PVY resistance by inoculation test; 13 transgenic plants including 7 for 3'-deleted Nlb, 3 for 5'-deleted Nlb, and 3 for ADD-mutant Nlb appeared to be resistant at 4 weeks after inoculation with PVY-VN. Among the 13 transgenic tobacco plants, 8 plants had no symptom up to 14 weeks after inoculation. The progenies ($T_1$) from self-fertilization of the transgenic lines varied 0.0% to 81.2% in their resistance (% of resistant plants). The analysis of Nlb-31deleted, -5'deleted and -ADD mutant in the $T_1$ plants by polymerase chain reaction (PCR) showed that Nlb-3'deleted, -5'deleted and -ADD mutants were detected in all of the resistant plants. These results suggest that the PVY resistance was inherited in the $T_1$ generation.

• Journal of Pharmaceutical Investigation
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• 제33권4호
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• pp.267-272
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• 2003

Herpes simplex virus (HSV) thymidine kinase (TK) has been widely used for suicidal gene therapy in combination with nucleoside analogs such as ganciclovir (GCV). The use of HSV-TK is limited due to the side effect of GCV at high concentrations. Previous studies showed that stable transfectants of mutant HSV-TK with enhanced affinity to GCV were killed at lower GCV concentrations. In this study, we tested whether mutant HSV-TK can provide enhanced suicidal effect when transiently transfected with Epstein-Barr virus (EBV)-based plasmid. EBV-based plasmid which contains OriP and EBNA-1 sequence is well known for a stable episomal maintenance in human cells. Optimal transfection condition was assessed for SNU-638 gastric cancer cell line using polyetylnimine (PEI). Maximum transfection efficiency was achieved when DNA:PEI was 1:3 (w/v). Cytotoxicities of mutant and wild type HSV-TK were compared before and after partially selecting transfected cells. The cells were sensitive to $100\;{\mu}g/ml$ hygromycin. Following GCV treatment, more cells were killed after hygromycin selection than before selection. The mutant HSV-TK showed enhanced cytotoxicity compared with the wild type HSV-TK. Our results suggest that the EBV-based plasmid encoding mutant HSV-TK may be useful to treat the diseases caused by uncontrolled cell proliferation such as cancer and rheumatoid arthritis.

• 제어로봇시스템학회논문지
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• 제17권8호
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• pp.753-759
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• 2011

The HIV (Human Immunodeficiency Virus) causes AIDS (Acquired Immune Deficiency Syndrome). The process of infection and mutation by HIV can be described by a 3rd order state equation. For this HIV model that includes the dynamics of the mutant virus, we present a parameter estimation scheme using two state variables sporadically measured, out of the three, by employing a genetic algorithm. It is assumed that these non-uniformly sampled measurements are subject to random noises. The effectiveness of the proposed parameter estimation is demonstrated by simulations. In addition, the estimated parameters are used to analyze the equilibrium points of the HIV model, and the results are shown to be consistent with those previously obtained.

• BMB Reports
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• 제46권1호
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• pp.53-58
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• 2013

We constructed deletion mutants and seven point mutants by polymerase chain reaction to investigate the specificity of feline foamy virus integrase functional domains. Complementation reactions were performed for three enzymatic activities such as 3'-end processing, strand transfer, and disintegration. The complementation reactions with deletion mutants showed several activities for 3'-end processing and strand transfer. The conserved central domain and the combination of the N-terminal or C-terminal domains increased disintegration activity significantly. In the complementation reactions between deletion and point mutants, the combination between D107V and deletion mutants revealed 3'-end processing activities, but the combination with others did not have any activity, including strand transfer activities. Disintegration activity increased evenly, except the combination with glutamic acid 200. These results suggest that an intact central domain mediates enzymatic activities but fails to show these activities in the absence of the N-terminal or C-terminal domains.