• Title/Summary/Keyword: Oligonucleotide

Search Result 444, Processing Time 0.036 seconds

Rhodobacter capsulatus bc$_{1}$ 복합체의 일차 구조

  • 채영규
    • The Microorganisms and Industry
    • /
    • v.14 no.3
    • /
    • pp.7-11
    • /
    • 1988
  • 이글에서는 pet A, B, C에 대한 일차적인 아미노산 배열을 알았고 여기서 우리는 위의 정보들을 토대로하여 R. capsulatus에 대한 site-specific oligonucleotide-directed mutagenesis 등 여러 연구를 할 수가 있을 것이다.

  • PDF

Effects of miR-155 Antisense Oligonucleotide on Breast Carcinoma Cell Line MDA-MB-157 and Implanted Tumors

  • Zheng, Shu-Rong;Guo, Gui-Long;Zhai, Qi;Zou, Zhang-Yong;Zhang, Wei
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.4
    • /
    • pp.2361-2366
    • /
    • 2013
  • Diverse studies have shown that miR-155 is overexpressed in different tumor types. However, the precise molecular mechanism of the ectopic expression of miR-155 in breast cancer is still poorly understood. To further explore the role of miR-155 in breast tumorigenesis, we here assessed the influence of miR-155 antisense oligonucleotide (miR-155 ASO) on MDA-MB-157 cell viability and apoptosis in vitro. Furthermore, the effects of inhibitory effects of miR-155 on the growth of xenograft tumors in vivo were determined with performance of immunohistochemistry to detect expression of caspase-3, a pivotal apoptosis regulatory factor, in xenografts. Transfection efficiency detected by laser confocal microscope was higher than 80%. The level of miR-155 expression was significantly decreased (P<0.05) in the cells transfected with miR-155 ASO, compared with that in cells transfected with a negative control. After being transfected with miR-155 ASO, the viability of MDA-MB-157 cells was reduced greatly (P<0.05) and the number of apoptotic cells was increased significantly. Additionally, miR-155 ASO inhibited the growth of transplanted tumor in vivo and significantly increased the expression of caspase-3. Taken together, our study revealed that miR-155 ASO can induce cell apoptosis and inhibit cell proliferation in vitro. Moreover, miR-155 ASO could significantly repress tumor growth in vivo, presumably by inducing apoptosis via caspase-3 up-regulation. These findings provide experimental evidence for using miR-155 as a therapeutic target of breast carcinoma.

Modulation of Antibody Responses against Gnathostoma spinigerum in Mice Immunized with Crude Antigen Formulated in CpG Oligonucleotide and Montanide ISA720

  • Intapan, Pewpan M.;Hirunpetcharat, Chakrit;Kularbkaew, Churairat;Yutanawiboonchai, Wiboonchai;Janwan, Penchom;Maleewong, Wanchai
    • Parasites, Hosts and Diseases
    • /
    • v.51 no.6
    • /
    • pp.637-644
    • /
    • 2013
  • This study aimed to investigate the antibody responses in mice immunized with Gnathostoma spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. spinigerum infection potently induces a Th2-type biased response.

Design, Optimization and Verification of 16S rRNA Oligonucleotide Probes of Fluorescence in-situ Hybridization for Targeting Clostridium spp. and Clostridium kluyveri

  • Hu, Lintao;Huang, Jun;Li, Hui;Jin, Yao;Wu, Chongde;Zhou, Rongqing
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.11
    • /
    • pp.1823-1833
    • /
    • 2018
  • Fluorescence in-situ hybridization (FISH) is a common and popular method used to investigate microbial communities in natural and engineered environments. In this study, two specific 16S rRNA-targeted oligonucleotide probes, CLZ and KCLZ, were designed and verified to quantify the genus Clostridium and the species Clostridium kluyveri. The optimal concentration of hybridization buffer solution for both probes was 30% (w/v). The specificity of the designed probes was high due to the use of pellets from pure reference strains. Feasibility was tested using samples of Chinese liquor from the famed Luzhou manufacturing cellar. The effectiveness of detecting target cells appears to vary widely in different environments. In pit mud, the detection effectiveness of the target cell by probes CLZ and KCLZ was 49.11% and 32.14%, respectively. Quantitative analysis by FISH technique of microbes in pit mud and fermented grains showed consistency with the results detected by qPCR and PCR-DGGE techniques, which showed that the probes CLZ and KCLZ were suitable to analyze the biomass of Clostridium spp. and C. kluyveri during liquor fermentation. Therefore, this study provides a method for quantitative analysis of Clostridium spp. and C. kluyveri and monitoring their community dynamics in microecosystems.

Identification of Viral RNA by Light Microscopic in situ Hybridization (광학현미경 In Situ Hybridization에 의한 Viral RNA 증명)

  • 최원기;주경웅;김석홍
    • Biomedical Science Letters
    • /
    • v.2 no.2
    • /
    • pp.249-255
    • /
    • 1996
  • In this paper, a in situ hybridization(ISH) has been used to investigate the yield of viral RNA expression from each organ tissues. It is studied to establish a rapidly, specific diagnostic method detecting rabbit haemorrhagic disease virus(RHDV) RNA in 10% formalin-fixed, paraffin-em-bedded tissues of naturally RHDV-infected rabbits using oligonucleotide probe to be made by RHDV total sequences. Biotin was used as the oligonucleotide probe marker. in situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method. All ISH procedure of RHDV were completed to Mi-croProbe$^{TM}$ capillary action system within 1-2 hours. In this report, RHDV was distributed widely in the cytoplasm of liver cell and the cortex of kidney but lung tissue and medulla of kidney were showed to positive reaction at locally. Although not entirely free of technical limitations, nucleic acid identification holds advantages over other diagnostic tests, including exquisite sensitivity, specificity, interchangeability and speed. It is expected that, in the immediate future viral nucleic acid detection will be a prominent part of the methods used in histopathology.

  • PDF

Development of the Large-Scale Oligonucleotide Chip for the Diagnosis of Plant Viruses and its Practical Use

  • Nam, Moon;Kim, Jeong-Seon;Lim, Seungmo;Park, Chung Youl;Kim, Jeong-Gyu;Choi, Hong-Soo;Lim, Hyoun-Sub;Moon, Jae Sun;Lee, Su-Heon
    • The Plant Pathology Journal
    • /
    • v.30 no.1
    • /
    • pp.51-57
    • /
    • 2014
  • A large-scale oligonucleotide (LSON) chip was developed for the detection of the plant viruses with known genetic information. The LSON chip contains two sets of 3,978 probes for 538 species of targets including plant viruses, satellite RNAs and viroids. A hundred forty thousand probes, consisting of isolate-, species- and genus-specific probes respectively, are designed from 20,000 of independent nucleotide sequence of plant viruses. Based on the economic importance, the amount of genome information, and the number of strains and/or isolates, one to fifty-one probes for each target virus are selected and spotted on the chip. The standard and field samples for the analysis of the LSON chip have been prepared and tested by RT-PCR. The probe's specific and/or nonspecific reaction patterns by LSON chip allow us to diagnose the unidentified viruses. Thus, the LSON chip in this study could be highly useful for the detection of unexpected plant viruses, the monitoring of emerging viruses and the fluctuation of the population of major viruses in each plant.

Cloning of the non-virion (NV) of a Korean Isolate of Infectious Hematopoietic Necrosis and Identification of the Role of the NV in IHNV Replication (한국에서 분리된 전염성 조혈괴저 바이러스의 non-virion (NV) 단백질의 유전자 클로닝 및 바이러스 증식에서의 역할)

  • 문창훈;조화자;윤원준;박정재;박정민;김현주;도정완;이주양;임채렬
    • Korean Journal of Microbiology
    • /
    • v.36 no.2
    • /
    • pp.103-108
    • /
    • 2000
  • We have cloned and analyzed cDNA coding for non-virion (NV) protein of the m V - P R T The NV gene contained 336 bp open readmg frame and encoded a protein of 11 1 amino acids with a molecular weight of 13.2 kDa. The deduced amino acid sequence of NV of IHNVPRT was found to be 90-95% identical to those of foreign isolates of IHNV. These results indicate that NV gene of the MNV is highly conserved among &ifferent strains of THNV Northern blot analyses revealed that the levels of NV gene expression were strongly elevated after 20 h post-infection. In order to identify the role of NV in the replication of MNV in fish cells, IHNVinfected cells were treated with antisense oligonucleotides. While IHNV-PRT exposed to glycoprotein (G) antisense oligonucleotide showed severely reduced growth, the growth of virus exposed to NV antisense oligonucleotide was not affected by NV antisense oligonucleotide, which suggests that NV is not essential for replication of IHNV in fish cells.

  • PDF