• Title/Summary/Keyword: Complementary Sequence

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Structural Study of Antisense Dimers, Modified Adenosine-Thymidine Phosphorothioate

  • Jung, Kyeong-Eun;Yang, Mi-Rim;Lee, Kwang-Jun;Lim, Hong;Jung, Ji-Hyun;Lim, Yoon-Gho;Cho, Youl-Hee;Shin, Dong-Hoon;Lee, Chul-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.889-892
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    • 2000
  • Antisense molecules are structurally simple linear oligomers of nucleotides. They can recognize a complementary sequence by base pairing, therefore, antisense drugs composed of 15-16 bases are potentially useful, unlike drugs such as protein agonists, antagonists, and inhibitors. Since antisense oligomers are classified as nucleotides, they are subject to attack by nucleases. In order to be antisense drugs resistant to degradation by nucleases, the structural modifications in the linkages, bases, and sugars to satisfy this requirement are considerable. We attempted in this study, to synthesize 16-mer antisenses with a modified linkage and adenosine. When studying on the three-dimensional structure of the oligomer, however, the existence of isomers may complicate the interpretation of the NMR data. Therefore, an attempt was made to eliminate the above problem, thus, two dimers were synthesized and their structural studies were carried out.

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Application of Geomagnetic Field-Based Indoor Positioning Technology in the Formwork Stage (거푸집공사 단계에서의 지구자기장 기반 작업자 실내측위기술 적용 방법)

  • Kim, Hyungjun;Lee, Changwoo;Kim, Hyeonmin;Ahn, Heejae;Lee, Changsu;Cho, Hunhee
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2023.05a
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    • pp.213-214
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    • 2023
  • Positioning information of workers is important for safety management at construction sites. Among the various indoor positioning technologies, geomagnetic fields-based technology is more economical and has less error than other technologies. However, there is a problem that the installation and dismantling of materials such as formwork at construction sites can cause degradation in positioning performance. Therefore, in this study, the distortion of the geomagnetic field near euro-form was quantitatively measured and the application method of geomagnetic field-based indoor positioning technology on formwork stage was presented. The results showed that the distortion occurred within 10cm of the wall and column form, but positioning accuracy could be affected up to 60cm from the form due to the characteristic of geomagnetic field-collecting technology. Therefore, applying this technology to the formwork stage requires complementary measures, such as using other positioning techniques up to 60 cm near the formwork, or excluding distorted area when positioning. It is expected that this study can contribute to the efficient safety management of workers by suggesting ways to prevent an increase in positioning error when applying geomagnetic field sequence-based indoor positioning technology during the formwork stage.

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Association of A/T Rich Microsatellites with Responses to Artificial Selection for Larval Developmental Duration in the Silkworm Bombyx mori

  • Pradeep, Appukuttan Nair Retnabhavan;Awasthi, Arvind Kumar;Urs, Raje Siddaraje
    • Molecules and Cells
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    • v.25 no.4
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    • pp.467-478
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    • 2008
  • Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) ($29.428{\pm}0.723days$) and shorter larval duration (SLD) ($22.573{\pm}0.839days$) lines from a base, inbred population of Nistari (larval span of $23.143{\pm}0.35days$). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 ($CA_7$) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the $F_2$ generation of the $LLD{\times}SLD$ lines, the alleles of Bmsat106 and $UBC873_{1074bp}$ segregated in a 1:1 ratio and the loci were present only in the LLD individuals. $UBC873_{823bp}$ was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the $UBC873_{1074bp}$ sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.

Identification of copy number variations using high density whole-genome single nucleotide polymorphism markers in Chinese Dongxiang spotted pigs

  • Wang, Chengbin;Chen, Hao;Wang, Xiaopeng;Wu, Zhongping;Liu, Weiwei;Guo, Yuanmei;Ren, Jun;Ding, Nengshui
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.12
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    • pp.1809-1815
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    • 2019
  • Objective: Copy number variations (CNVs) are a major source of genetic diversity complementary to single nucleotide polymorphism (SNP) in animals. The aim of the study was to perform a comprehensive genomic analysis of CNVs based on high density whole-genome SNP markers in Chinese Dongxiang spotted pigs. Methods: We used customized Affymetrix Axiom Pig1.4M array plates containing 1.4 million SNPs and the PennCNV algorithm to identify porcine CNVs on autosomes in Chinese Dongxiang spotted pigs. Then, the next generation sequence data was used to confirm the detected CNVs. Next, functional analysis was performed for gene contents in copy number variation regions (CNVRs). In addition, we compared the identified CNVRs with those reported ones and quantitative trait loci (QTL) in the pig QTL database. Results: We identified 871 putative CNVs belonging to 2,221 CNVRs on 17 autosomes. We further discarded CNVRs that were detected only in one individual, leaving us 166 CNVRs in total. The 166 CNVRs ranged from 2.89 kb to 617.53 kb with a mean value of 93.65 kb and a genome coverage of 15.55 Mb, corresponding to 0.58% of the pig genome. A total of 119 (71.69%) of the identified CNVRs were confirmed by next generation sequence data. Moreover, functional annotation showed that these CNVRs are involved in a variety of molecular functions. More than half (56.63%) of the CNVRs (n = 94) have been reported in previous studies, while 72 CNVRs are reported for the first time. In addition, 162 (97.59%) CNVRs were found to overlap with 2,765 previously reported QTLs affecting 378 phenotypic traits. Conclusion: The findings improve the catalog of pig CNVs and provide insights and novel molecular markers for further genetic analyses of Chinese indigenous pigs.

Cloning and Characterization of the Paraquat Resistance-Related Genes from Ochrobactrum anthropi JW-2 (Ochrobactrum anthropi JW-2 유래의 Paraquat 내성유전자 PqrA의 주변 유전자군 분석)

  • Bae Eun-Kyung;Lee Hyo-Shin;Won Sung-Hye;Lee Byung-Hyun
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.15-22
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    • 2006
  • A 4,971 bp chromosomal DNA fragment containing the pqrA, paraquat resistance gene, was cloned from Ochrobactrum anthropi JW-2, and the complete nucleotide sequence was determined. Nucleotide and deduced amino acid sequences of the fragment revealed the presence of 4 complete ORFs (orf2, pqrA, orf3, orf4) and two incomplete ORFs(orf1, orf5). Orf1, pqrA, orf4 and orf5 exists at the direct strand but orf2 and orf3 exists at the reverse complementary strand. Orf1 which of incomplete sequences without start codon shares homology with ATP binding region of the response regulator receiver. Orf2 shares high homology with members of the tetR family of transcriptional repressor which have a helix-turn-helix (H-T-H) motif. Therefore, the orf2 is predicted as a transcriptional repressor of pqrA and is designated as pqrR2. Orf3 shares high homology with the members of the lysR family acting as a transcriptional activator which have both of a H-T-H motif at the N-terminal region and substrate binding domain at the C-terminal region. Therefore, the orf3 is predicted as a transcriptional activator of pqrA and is designated as pqrR1. Orf4 shows homology with the periplasmic substrate-binding protein of amino acid ABC transporter. Orf5 which of incomplete sequences without stop codon revealed the homology with the permeases protein of amino acid ABC transporter.

Susceptibility-Weighted Imaging as a Distinctive Imaging Technique for Providing Complementary Information for Precise Diagnosis of Neurologic Disorder (신경계 질환에 관한 정확한 진단을 위해 다양한 보완 정보를 제공하는 독특한 영상 기법으로서의 자기화율 강조 영상)

  • Byeong-Uk Jeon;In Kyu Yu;Tae Kun Kim;Ha Youn Kim;Seungbae Hwang
    • Journal of the Korean Society of Radiology
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    • v.82 no.1
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    • pp.99-115
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    • 2021
  • Various sequences have been developed for MRI to aid in the radiologic diagnosis. Among the various MR sequences, susceptibility-weighted imaging (SWI) is a high-spatial-resolution, three-dimensional gradient-echo MR sequence, which is very sensitive in detecting deoxyhemoglobin, ferritin, hemosiderin, and bone minerals through local magnetic field distortion. In this regard, SWI has been used for the diagnosis and treatment of various neurologic disorders, and the improved image quality has enabled to acquire more useful information for radiologists. Here, we explain the principle of various signals on SWI arising in neurological disorders and provide a retrospective review of many cases of clinically or pathologically proven disease or components with distinctive imaging features of various neurological diseases. Additionally, we outline a short and condensed overview of principles of SWI in relation to neurological disorders and describe various cases with characteristic imaging features on SWI. There are many different types diseases involving the brain parenchyma, and they have distinct SWI features. SWI is an effective imaging tool that provides complementary information for the diagnosis of various diseases.

Photo-controlled gene expression by fluorescein-labeled antisense oligonucleotides in combination with visible light irradiation

  • Ito, Atsushi;Kaneko, Tadashi;Miyamoto, Yuka;Ishii, Keiichiro;Fujita, Hitoshi;Hayashi, Tomonori;Sasaki, Masako
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.451-453
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    • 2002
  • A new concept of "photo" -antisense method has been evaluated, where the inhibition of gene expression by the conventional antisense method is enhanced by photochemical binding between antisense oligonucleotides conjugated with photo-reactive compound and target mRNA or DNA. Fluorescein labeled oligodeoxyribonucleotides (F-DNA) was delivered to cell nuclei in the encapsulated form in multilamellar lecithin liposomes with neutral charge. F-DNA was previously shown to photo-bind to the complementary stranded DNA, and the delivery system using neutral liposome to be effective in normal human keratinocytes. In the present study, we used human kidney cancer G401.2/6TG.1 cell line to be advantageous in reproducible experiments. p53 was adopted as a target gene since antisense sequence information has been accumulated. The nuclear localization ofF-DNA was identified by comparing the fluorescence ofF-DNA with that of Hoechst 33258 under fluorescence microscope. After 7hr incubation to accumulate p53 protein induced by UV -B, p53 protein was quantified by Western blot. After 2hrs from F-DNA application, about 30% of cell population incorporated F-DNA in their nuclei with some morphological change possibly due to liposomal toxicity. Irradiation of visible light longer than 400nm from solar simulator at this time enhanced the inhibitory action of antisense F-DNA. The present results suggest that photo-antisense method is promising to control gene expression in time and space dependent manner. Further improvement of F-DNA delivery to cancer cells in the stability and toxicity is in progress. progress.

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Development of Enrichment Semi-nested PCR for Clostridium botulinum types A, B, E, and F and Its Application to Korean Environmental Samples

  • Shin, Na-Ri;Yoon, So-Yeon;Shin, Ji-Hun;Kim, Yun Jeong;Rhie, Gi-eun;Kim, Bong Su;Seong, Won Keun;Oh, Hee-Bok
    • Molecules and Cells
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    • v.24 no.3
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    • pp.329-337
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    • 2007
  • An enrichment semi-nested PCR procedure was developed for detection of Clostridium botulinum types A, B, E, and F. It was applied to sediment samples to examine the prevalence of C. botulinum in the Korean environment. The first pair of primers for the semi-nested PCR was designed using a region shared by the types A, B, E, and F neurotoxin gene sequences, and the second round employed four nested primers complementary to the BoNT/A, /B, /E, and /F encoding genes for simultaneous detection of the four serotypes. Positive results were obtained from the PCR analysis of five of 44 sediments (11%) collected from Yeong-am Lake in Korea; all were identified as deriving from type B neurotoxin (bontb) genes. Two of the C. botulinum type B organisms were isolated, and their bontb genes sequenced. The deduced amino acid sequences of BoNT/B showed 99.5 and 99.8% identity with the amino acid sequence of accession no. AB084152. Our data suggest that semi-nested PCR is a useful tool for detecting C. botulinum in sediments, and renders it practicable to conduct environmental surveys.

Isolation and Characterization of Watermelon Isolate of Cucumber green mottle mosaic virus(CGMMV-HY1) from Watermelon Plants with Severe Mottle Mosaic Symptoms

  • Shim, Chang-Ki;Han, Ki-Soo;Lee, Jung-Han;Bae, Dong-Won;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.21 no.2
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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.

Analysis of the Type, Function, and Structure of Inscriptions in Middle School Science Textbooks: Focus on Earth Science Content of the 7th National Curriculum (중학교 과학 교과서에 사용된 시각자료의 유형, 기능 및 구조 분석: 제7차 교육과정 지구과학 내용을 중심으로)

  • Lee, Ki-Young
    • Journal of the Korean earth science society
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    • v.30 no.7
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    • pp.897-908
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    • 2009
  • Inscriptions are integral parts of the current science textbooks, and become medium that makes sense of scientific concepts effectively. This study analyzed the inscriptions from five science textbooks at the secondary level in terms of its type, function, and structures. Analyzing types of inscriptions resulted that the ratio of frequently used photographs and illustrations were much higher than that of graphs and tables used. In case of function analysis, illustrative and complementary functions were dominant in terms of frequency and ratio, whereas explanatory, decorative and inquisitive functions were relatively rare. Inscriptions of single structure was much more than multiple structure. Multiple structure of pair and series were mainly used for representing contrast and sequence, respectively. The proportion of inscriptions in type, function and structure across the textbooks was significantly different. Based on the results, it was suggested that semiotic study about the function and the structure of inscriptions need to be carried out in earth science area to better understand the pedagogical implications of inscriptions in school science.