• 제목/요약/키워드: viral DNA

검색결과 443건 처리시간 0.021초

Perspectives of AIDS Vaccine Development: T Cell-based Vaccine

  • Sung, Young Chul
    • IMMUNE NETWORK
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    • 제2권1호
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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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    • 제27권7호
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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.

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

  • 이평우;김윤철;백우현
    • 대한바이러스학회지
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    • 제26권1호
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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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B형 및 C형 간염에 대한 한약 치료의 효과 - 체계적 고찰 연구 (Herbal Medicine for the Treatment of Viral Hepatitis B and C: A Systematic Review of Randomized Controlled Trials)

  • 김승모;이유리;조나경;최홍식;김경순
    • 대한한방내과학회지
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    • 제42권4호
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    • pp.455-474
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    • 2021
  • 본 연구는 B형 및 C형 간염에 대한 한약 치료의 효과를 평가하기 위해 무작위 임상연구를 대상으로 체계적 문헌고찰 및 메타분석을 시행하였다. 검색엔진은 EMBASE, Pubmed, NDSL, KMBASE, KISS, KISTI, Koreamed, Koreantk, Oasis database를 이용하였으며 국내의 검색 엔진 키워드는 '간염' 또는 '바이러스성간염', '한약', '무작위 배정 임상시험'을, 국외 검색 엔진 키워드는 'hepatitis' or 'viral hepatitis' and 'herbal medicine' or 'traditional chinese medicine' and 'randomized controlled trial'을 이용하였다. 연구 결과 15개의 무작위배정 임상시험을 선택되었으며, 그 중 통계적으로 유의미한 결과를 나타낸 연구는 한약과 양약 복합치료군이 양약을 투여한 대조군에 비해 HBV DNA loss에서 높은 효과를 보였다. 한약과 양약 복합치료군이 양약을 투여한 대조군에 비해 HBeAg loss에서 통계적으로 유의한 효과를 보였으며, 한약과 양약 복합치료군이 양약을 투여한 대조군에 비해 ALT가 감소하였다. 본 연구는 B형 및 C형 간염환자를 대상으로 한약의 효과를 확인하기 위해 기존에 시행된 RCT 연구를 대상으로 체계적 문헌고찰을 시행하였으며, 한약과 양약의 복합치료가 양약 단독치료에 비해 치료효과가 더 높은 것으로 나타났다.

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

  • 손혜정;김진석
    • 폴리머
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    • 제29권1호
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    • pp.69-73
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    • 2005
  • 비바이러스성 유전자 전달체의 주요 단점인 낮은 transfection 효율에 기인한 반복투여 등을 극복하기 위하여 poly (D,L-lactide-co-glycolide)를 이용하여 DNA가 봉입된 미립구를 제조하였다. pDNA 그 자체 또는 여러 비율의 키토산/pDNA 복합체를 사용하여 봉입하였고, 그 결과 44%(pDNA 그 자체), 5%(0.7:1 미토산/pDNA 복합체), 그리고 8%(1:1 키토산/pDNA 복합체)의 봉입효율을 나타내었다. 주사전자현미경(SEM)을 통해 본 표면구조에서는 미립구 제조 직후에서는 매우 매끈한 구형을 보이다가 제조 후 41일 경에는 찌그러진 다공성의 구조를 보였는데 이는 미립구 제조에 사용한 poly(D,L-lactic-co-glycolic acid)(PLGA) 고분자의 분해에 의한 것으로 생각된다. 시험관내 방출실험에서는 0.7:1 키토산/pDNA 복합체를 사용한 미립구에서 47%의 pDNA가 26일만에 방출된데 반해, pDNA 그 자체 혹은 1:1 키토산/pDNA 복합체를 사용한 미립구에서는 각각 15% 혹은 32%의 pDNA 방출을 나타내었다.

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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    • 제45권3호
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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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    • 제11권3호
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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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파밤나방 핵다각체병 바이러스의 생화학적 특성 (Biochemical Characteristics of Spodoptera exigua Nuclear Polyhedrosis Virus)

  • 진병래;박범석;재연호;강석권
    • 한국응용곤충학회지
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    • 제30권2호
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    • pp.144-149
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    • 1991
  • 국내에서 분리된 파밤나방 핵다각체병바이러스(Spodoptera exigua nuclear polyhedrosis virus: SeNPV)의 생화학적인 특성을 규명하기 위하여 몇가지 실험을 행하였다. SeNPV는 하나의 envelopeso내에 다수의 nucleocapsid가 존재하는 MNPV(multiple embeded NPV)형태였다. 다각체단백질은 분자량 30kb의 단일 band로 나타났으며, Spodoptera litura NPV와 Bombyx mori NPV의 다각체단백질 항체에 반응하여 뚜렷한 침강선을 형성하였다. 비리온 단백질을 은염색한 결과, 많은 수의 minor band들이 포함된 49개의 band로 나타났으며, 바이러스 DNA를 분리하여 여러종의 제한효소에 의한 대략적인 genome size는 약 110kb 였다.

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

  • Young Ho Kim;Eun Kyung Park;Soon Yong Chae;Sang Seock Kim;Kyung-Hee Paek;Hye Sun Cho
    • 한국연초학회지
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    • 제20권1호
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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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