• Molecules and Cells
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• 제40권5호
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• pp.339-345
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• 2017

Retroviral and lentiviral vectors are mostly pseudotyped and often purified and concentrated via ultracentrifugation. In this study, we quantified and compared the stabilities of retroviral [murine leukemia virus (MLV)-based] and lentiviral [human immunodeficiency virus (HIV)-1-based] vectors pseudotyped with relatively mechanically stable envelope proteins, vesicular stomatitis virus glycoproteins (VSVGs), and the influenza virus WSN strain envelope proteins against ultracentrifugation. Lentiviral genomic and functional particles were more stable than the corresponding retroviral particles against ultracentrifugation when pseudotyped with VSVGs. However, both retroviral and lentiviral particles were unstable when pseudotyped with the influenza virus WSN strain envelope proteins. Therefore, the stabilities of pseudotyped retroviral and lentiviral vectors against ultracentrifugation process are a function of not only the type of envelope proteins, but also the type of viral internal core (MLV or HIV-1 core). In addition, the fraction of functional viral particles among genomic viral particles greatly varied at times during packaging, depending on the type of envelope proteins used for pseudotyping and the viral internal core.

• 미생물학회지
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• 제31권3호
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• pp.184-188
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• 1993

Novel Ustilagomaydis strains, designated as SH1 to 14 containing new types of ds RNA segments, are identified from corn smut in Korea. Among 14 isolates, 7 isolates appear to posses virus particles and the other isolates may contain dsRNA as a plasmid form. The pattern of dsRNA is highly diverse form a typical P-type containing one or more of H, M, and L dsRNAs to the one containing one or move M dsRNAs. It is likely that the strains containing H dsRNA posses virus particles which were confirmed by sucrose density gradient followed with different range of specificity and the activity of the strain (SH14) is stronger than A4 toxin. The sensitivity of 14 isolates is also very diverse and two strains (SH10, SH11) appear tobe universal sensitve strains against 5 tested toxin samples.

• 농업생명과학연구
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• 제44권6호
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• pp.141-146
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• 2010

본 연구에서는 국내에서 식용으로 재배되고 있는 팽이버섯의 균사체에 대하여 Agrobacterium을 이용한 형질전환을 시도하였다. 특히 물리적 연마제인 미세 aluminum oxide 입자를 팽이 균사체와 함께 강하게 교반함으로써 물리적 상해를 지니는 균사체를 제조하였으며 감압침윤에 의한 Agrobacterium 이용 형질전환을 시도하였다. Hygromycin 저항성을 이용한 선발 결과, 대조군에서는 균사체 생육이 전혀 관찰되지 않은 반면 물리적 상해군에서는 형질전환균사체 생육이 확인되었다. Genomic DNA PCR에 의한 유전자 도입을 확인함으로써 팽이균사체에 대한 매우 간편한 형질전환법을 제시할 수 있었다.

• 한국축산식품학회지
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• 제40권5호
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• pp.746-757
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• 2020

Enterotoxigenic Escherichia coli (ETEC) is the major pathogenic E. coli that causes diarrhea and edema in post-weaning piglets. In this study, we describe the morphology and characteristics of ØCJ19, a bacteriophage that infects ETEC, and performed genetic analysis. Phage ØCJ19 belongs to the family Myoviridae. One-step growth curve showed a latent phase of 5 min and burst size of approximately 20 phage particles/infected cell. Phage infectivity was stable for 2 h between 4℃ and 55℃, and the phage was stable between pH 3 and 11. Genetic analysis revealed that phage ØCJ19 has a total of 49,567 bases and 79 open reading frames (ORFs). The full genomic sequence of phage ØCJ19 showed the most similarity to an Escherichia phage, vB_EcoS_ESCO41. There were no genes encoding lysogeny, toxins, virulence factors, or antibiotic resistance in this phage, suggesting that this phage can be used safely as a biological agent to control ETEC. Comparative genomic analysis in terms of the tail fiber proteins could provide genetic insight into host recognition and the relationship with other coliphages. These results showed the possibility to improve food safety by applying phage ØCJ19 to foods of animal origin contaminated with ETEC and suggests that it could be the basis for establishing a safety management system in the animal husbandry.

• Journal of Applied Biological Chemistry
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• 제58권3호
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• pp.273-279
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• 2015

해양 미세조류인 Phaeodactylum tricornutum은 게놈 염기서열이 완전히 밝혀진 규조류로서, 형질전환 방법이 개발되어 있고, 여러 가지 분자생물학적 연구 기술이 개발되어 규조류 연구에서 모델 종으로 여겨지고 있다. 본 연구의 목적은 methylerythritol phosphate (MEP) 대사경로의 마지막 효소인 (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR)를 코딩하는 P. tricornutum의 Pthdr 유전자를 P. tricornutum에 도입하여 형질전환체를 확보하는 것이다. 유전자 도입 방법은 gold microcarrier를 사용한 bombardment 방법을 사용하였고, 형질전환 유무 및 목적 유전자의 전사체 확인에는 각각 genomic DNA-PCR 및 cDNA-PCR 방법을 사용하였다. 양성대조군으로 egfp 유전자를 P. tricornutum에 도입하여 최종적으로 eGFP 단백질이 발현되는 것을 형광 공초점 현미경을 통해 확인하였다. 이를 바탕으로, 확보된 Pthdr 형질전환체에서도 도입한 Pthdr 유전자로부터 발현된 PtHDR 효소도 잘 발현될 것으로 추측할 수 있었다. 이렇게 준비된 Pthdr 형질전환체는 추후 연구를 통해, P. tricornum의 유용물질인 카로티노이드의 생합성 과정 연구 및 고부가가치 카로티노이드 과발현 균주 개발 등에 유용한 정보를 제공할 것으로 기대된다.

• The Plant Pathology Journal
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• 제21권2호
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• pp.167-171
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• 2005

We isolated the Cucumber green mottle mosaic virus(CGMMV) particles from watermelon leaves and designated as CGMMV-HY1 as a watermelon isolate and attempted to characterize the pathogenic isolate responsible for such an epidemic in watermelon and also to monitor dominant viral isolates in greenhouse. The watermelon plants infected with CGMMV generally showed mottle mosaic, mosaic, growth stunting, necrosis and deformed fruit. The reactions of indicator plants to CGMMV-HY1 were the local lesions on Nicotiana tabacum cv. White Burley, Nicotiana tabacum cv. Samsun, and Chenopodium amaranticola, and the mosaic symptoms only on Cucumis sativus, but the CGMMV-HY1 did not infect Nicotiana sylvesytis, Datura stramonium, Chenopodium quinoa, and Petunia hybrida. Purified virus particles were rod-shaped and about 300 nm long. The coat protein (CP) of purified CGMMV-HY1 was single band with molecular weight of about 16.5 kDa which was confirmed by western blot analysis probed with monoclonal antibody of CGMMV-HY1. The genomic and subgenomic RNAs of 6.4 kb and 0.75 kb were revealed by the electrophoresis on 1.2% formaldehydedenatured agarose gel. Viral and complementary CGMMV-specific primer sets were designed for spanning the genome using previously reported CGMMV sequences. A 464bp of CP gene of CGMMV-HY1 was amplified by RT-PCR and cloned into PGEM-T easy vector. The nucleotide sequence of CP gene of CGMMV-HY1 shared 98%, 99%, and 100% identities with that of CGMMV strains W, KOM, and KW respectively. Based on these results, we identified CGMMV-HY1 as a CGMMV isolate of watermelon, a member of Tobamovirus.

• 대한수의학회지
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• 제32권1호
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• pp.77-82
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• 1992

세포 비병원성 소 설사병 바이러스가 감염된 우태아 폐세포를 이용 바이러스를 순수분리하여 이화학적 성상을 검사하였던 바 다음의 결과를 얻었다. 바이러스의 비중은 $1.090{\sim}1,114gm/cm^3$로 검출되었으며 $1,098gm/cm^3$에서 최대 감염가를 나타내었다. 면역 전자현미경을 이용한 형태학적 분석결과 30~80nm의 바이러스성 입자를 관찰할 수 있었으며 유전인자의 추출 분석결과 병원성 바이러스와의 전기영동상 유전자 규모에 있어서 차이는 인정되지 않았다. 그러나 비세포 병원성 소 설사병 바이러스는 병원성 바이러스에 비하여 세포외 배출이 적었으며 세포내에서 보다 많이 존재하는 것으로 관찰되었다.

• The Plant Pathology Journal
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• 제20권4호
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• pp.297-301
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• 2004

A Tomato spotted wilt virus (TSWV-KP) was isolated from Paprika (Capsicum annuum var. grossum) showing necrosis spot on the leaves and malformation of the fruit in Yesan, Korea. The virus infected Chenopodium amaranticolor, C. quinoa, Petunia hybrida, Nicotiana glutunosa, Gomphrena globosa, and Physalis floridana. Ten plants including tomato were observed to have systemic TWSV-KP infection. The virus produced necrosis or necrotic ring spots on the inoculated leaves and mosaic, vein necrosis or death on the upper leaves of Datura stramonium, N. clevarandii, N. rustica, and N.tabacum cvs. Thin sections of the infected leaf tissue contained spherical to oval particles, a characteristic of a Tospovirus. The virion contained three molecules of genomic RNAs, which were approximately 9.0, 4.9 and 3.0 kb. The nucleocapsid (N) protein of the purified virion migrated as a single band with molecular weight of about 29 kDa in SDS-PAGE. The N gene of TSWV-KP showed 96.5-97.2% and 97.7-98.5% identities to the three different TSWV isolates of Genbank Database at the nucleotide and amino acid, respectively.

• Journal of Applied Biological Chemistry
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• 제42권3호
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• pp.113-118
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• 1999

The transgenic plant of Citrus species (Citrus aurantium L.) was constructed with a fatty acid desaturase gene using microprojectile bombardment transformation system. The DNA of a fatty acid desaturase gene, fad7, constructed in pBI121 was coated onto tungsten particles ($1.1{\mu}m$) and introduced into callus cells by bombarding with 1100 psi of helium pressure, 1/4 in of gap distance, 7.0 cm of target distance and 27 in Hg of chamber vacuum. The bombarded cells were selected on the medium containing kanamycin. The selected cells were successfully regenerated into plantlets via somatic embryogenesis on the media containing plant growth regulators. The results of polymerase chain reaction analysis of genomic DNAs from the putative transformants showed that the introduced DNAs of fad7 were present in both the selected callus cells and the regenerated plantlets.

• BMB Reports
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• 제48권12호
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• pp.685-690
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• 2015

The eukaryotic genome is packed into chromatin, which is important for the genomic integrity and gene regulation. Chromatin structures are maintained through assembly and disassembly of nucleosomes catalyzed by histone chaperones. Asf1 (anti-silencing function 1) is a highly conserved histone chaperone that mediates histone transfer on/off DNA and promotes histone H3 lysine 56 acetylation at globular core domain of histone H3. To elucidate the role of Asf1 in the modulation of chromatin structure, we screened and identified small molecules that inhibit Asf1 and H3K56 acetylation without affecting other histone modifications. These pyrimidine-2,4,6-trione derivative molecules inhibited the nucleosome assembly mediated by Asf1 in vitro, and reduced the H3K56 acetylation in HeLa cells. Furthermore, production of HSV viral particles was reduced by these compounds. As Asf1 is implicated in genome integrity, cell proliferation, and cancer, current Asf1 inhibitor molecules may offer an opportunity for the therapeutic development for treatment of diseases.