• Title/Summary/Keyword: viral DNA

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Perspectives of AIDS Vaccine Development: T Cell-based Vaccine

  • Sung, Young Chul
    • IMMUNE NETWORK
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    • v.2 no.1
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    • pp.1-5
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    • 2002
  • Estimated number of adults and children newly infected with HIV-1 during 2001 alone is 5 million in total. An effective vaccine, in addition to education & public health approaches, has been believed to be the best option to stop the HIV-1 transmission, especially for developing countries. Among AIDS vaccine candidates, DNA vaccine is relatively safe and, in a certain extent, mimics some attributes of live attenuated vaccine, with regard to in vivo gene expression & the type of immunity induced. We recently demonstrated that DNA vaccines expressing SIVmac239 structural and regulatory genes, augmented with coadministration of IL-12 mutant induced the strongest T cell responses, resulting in low to undetectable setpoint viral loads, stable $CD4^+$ T cell counts, and no evidence of clinical diseases or mortality by day 420 after challenge. This finding is the second demonstration, following the protective result of live attenuated SIV vaccine in SIVmac-rhesus monkey model, which was known to have safety problem. So, our DNA vaccines could give a significant impact on HIV-1 epidemic by slowing or stopping the spread of HIV-1, leading to eventual eradication of HIV-1 and AIDS in the population.

Cloning and Characterization of Directly Amplified Antiviral Gene Interferon Alpha-2b (HulFN$\{alpha}$-2b) from Human Leukocytes Chromosomal DNA

  • Behravan, Javad;Ahmadpour, Hassan
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.776-780
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    • 2004
  • Interferons are cytokines that confer resistance to viral infection and inhibit cellular proliferation. The interferon alpha gene from human blood samples was amplified, cloned and expressed in E. coli (BL21). Leukocyte chromosomal DNA was used as a source of template DNA. Using specific primers, the gene for HulFN$\{alpha}$-2b was amplified and inserted into the E. coli vector, pET21b, by ligation of the HindIII and BamHI linkers of the vector and insert. The insert was further analyzed by PCR, DNA restriction mapping and sequencing, and expressed in a suitable E. coli strain. The production of this important cellular protein in the laboratory has significant applications in production of the recombinant pharmaceutical proteins.

Expression of Nucleocapsid Protein Gene of Maaji Virus and Use of the Protein as an Immunodiagnostic Antigen of Hemorrhagic Fever with Renal Syndrome (마지바이러스 Nucleocapsid Protein 유전자의 발현과 신증후 출혈열 진단용 항원으로의 이용)

  • Lee, Pyung-Woo;Kim, Yun-Cheol;Paik, Woo-Hyun
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.77-90
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    • 1996
  • Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.

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Herbal Medicine for the Treatment of Viral Hepatitis B and C: A Systematic Review of Randomized Controlled Trials (B형 및 C형 간염에 대한 한약 치료의 효과 - 체계적 고찰 연구)

  • Kim, Seung-mo;Lee, Yu-ri;Cho, Na-kyung;Choi, Hong-sik;Kim, Kyung-soon
    • The Journal of Internal Korean Medicine
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    • v.42 no.4
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    • pp.455-474
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    • 2021
  • The aim of this study was to perform a systematic review and meta-analysis of randomized controlled trials (RCTs) that applied herbal medicine to treat viral hepatitis B and C in order to determine the therapeutic efficacy of herbal medicine. EMBASE, Pubmed, NDSL, KMBASE, KISS, KISTI, Koreamed, Koreantk, and Oasis databases were searched to identify RCTs. The selected studies were assessed by the Cochrane group's risk of bias tool. A total of 15 RCTs were selected, and the hepatitis B virus (HBV) DNA reduction was significantly higher in patients treated with herbal medicine combined with Western medicine than in patients treated with herbal medicine. Herbal medicine combined with Western medicine was also superior to Western medicine alone in achieving hepatitis B e-antigen (HBeAg) and alanine aminotransferase [ALT] reduction. Only herbal medicine alone was not superior to Western medicine treatments in achieving HBV DNA, HBeAg, and ALT reduction.

Preparation and In Vitro Release of DNA-Loaded Poly(D,L-lactic-co-glycolic acid) Microspheres (DNA가 봉입된 Poly(D,L-lactic-co-glycolic acid) 미립구의 제조 및 시험관내 방출)

  • Son, Hye-Jung;Kim, Jin-Seok
    • Polymer(Korea)
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    • v.29 no.1
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    • pp.69-73
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    • 2005
  • To overcome the main disadvantages of non-viral gene delivery systems such as repeated administration due to the low transfection efficiency, poly(D,L-lactide-co-glycolide) was applied to encapsulate pDNA in its microsphere formulation. Free pDNA or various ratios (w/w) of chitosan/pDNA complexes was used for encapsulation, with the resulting encapsulation efficiency of 44%, 5%, and 8% for free pDNA, 0.7:1 and 1:1 ratios, respectively. Scanning electron micrographs of poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres encapsulating pDNA or chitosan-condensed pDNA revealed a smooth spherical shape immediately after microsphere preparation and a collapsed porous shape in 41 days due to the degradation of PLGA. In vitro release profile showed that the 0.7:1 (w/w) ratio formulation exerted 47% release in 26 days, whereas free pDNA or 1:1 (w/w) ratio formulation did only 15% or 32%, respectively.

Identification of anti-HIV and anti-Reverse Transcriptase activity from Tetracera scandens

  • Kwon, Hyeok-Sang;Park, Jung-Ae;Kim, Joo-Hwan;You, Ji-Chang
    • BMB Reports
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    • v.45 no.3
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    • pp.165-170
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    • 2012
  • We report here that an ethanol extract of Tetracera scandens, a Vietnamese medicinal plant, has anti-HIV activity and possesses strong inhibitory activity against HIV-1 reverse transcriptase (RTase). Using a MT-4 cell-based assay, we found that the T. scandens extract inhibited effectively HIV virus replication with an $IC_{50}$ value in the range of 2.0-2.5 ${\mu}g$/ml while the cellular toxicity value (CC50) was more than 40-50 ${\mu}g$/ml concentration, thus yielding a minimum specificity index of 20-fold. Moreover, the anti-HIV efficacy of the T. scandens extract was determined to be due, in part, to its potent inhibitory activity against HIV-1 RTase activity in vitro. The inhibitory activity against the RTase was further confirmed by probing viral cDNA production, an intermediate of viral reverse transcription, in virus-infected cells using quantitative DNA-PCR analysis. Thus, these results suggest that T. scandens can be a useful source for the isolation and development of new anti-HIV-1 inhibitor(s).

White Spot Syndrome Virus in Penaeid Shrimp Cultured in Korea

  • Shin, En-Joo;Park, Jae-Hak;Lee, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.394-398
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    • 2001
  • Because of the great concern over the possibility of contamination from the rod-shaped nuclear virus (PRDV) from Japan and white spot virus (WSSV) from Taiwan, most eggs used in Korean shrimp farms are currently obtained from local broodstock. In addition, the screening of imported broodstock for any viral presence at the National Fisheries Research and Development Institute is also mandatory. Nonetheless, massive mortality from white spot syndrome continues in Korea. In the present study, we present an improved PCR method to use tissue-extracted DNA instead of viral DNA extracted from a purified virus based on a sucrose density gradient, and produced results within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected only with PRDV, while Fenneropenaeus chinensis were infected with both PRDV and WSSV. In 1999, PRDV and WSSV were detected in F. chinensis with signs of infection, but not with WSSV alone.

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Biochemical Characteristics of Spodoptera exigua Nuclear Polyhedrosis Virus (파밤나방 핵다각체병 바이러스의 생화학적 특성)

  • 진병래;박범석;재연호;강석권
    • Korean journal of applied entomology
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    • v.30 no.2
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    • pp.144-149
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    • 1991
  • Biochemical characteristics of Spodoptera uigua nuclear polyhedrosis virus (SeNPV) isolated in Jinju were studied. SeNPV contained a number of nucleocapsids within a viral envelope embeded in polyhedra. The polyhedral protein of SeNPV was composed of a single polypeptide with a M. W. of 30kd. Double-immunodiffusion test showed that the polyhedral protein of SeNPV had common antigenic determinants with SINPV and BmNPV. Virion proteins of SeNPV were resolved into 49 polypeptides by silver staining after SDS-PAGE. The approximate genome size of SeNPV by restriction endonuclease analysis was 1l0kb.

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Resistance to Potato Virus Y Conferred by PVY Replicase Gene Sequence in Transgenic Burley Tobacco (감자바이러스 Y 복제 유전자로 형질전환된 버어리종 연초의 PVY에 대한 저항성 특성)

  • Young Ho Kim;Eun Kyung Park;Soon Yong Chae;Sang Seock Kim;Kyung-Hee Paek;Hye Sun Cho
    • Journal of the Korean Society of Tobacco Science
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    • v.20 no.1
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    • pp.50-56
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    • 1998
  • The complementary DNA (cDNA) of potato virus Y- vein necrosis strain (PVY-VN) replicase gene (Nlb) was transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Out of 25 putative transformants regenerated, 3 were resistant to PVY-VN, one highly resistant plant with no symptom until seed harvest time and the other two with mild chlorotic spot symptoms at late stages after infection. No symptom was observed in the highly resistant plant, while mild vein necrotic symptoms were developed on suckers of the moderately resistant plants after seed harvest time, In the first generation (T1) via self fertilization, resistance to susceptibility frequency in transgenic plants from the highly resistant transformant was about 3 : 1, while it was lowered much (about 1:2 and 1:19) in T1 of the moderately resistant transformants. In the second generation (T2) of the highly resistant plant, resistance frequencies were similar to T1, but resistance levels varied greatly and appeared to be decreased. Key words : potato virus Y, viral replicate gene, transgenic tobacco plants, resistance.

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