• 제목/요약/키워드: m-Sequence

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Specific Gene Silencing by Single Stranded Large Circular Antisense Molecules

  • Park, Jong-Gu
    • 대한의생명과학회지
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    • 제10권2호
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    • pp.65-73
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    • 2004
  • I report that single-stranded antisense as a part of large circular (LC-) genomic DNA of recombinant M13 phage exhibits enhanced stability, sequence specific antisense activity, and no need for target site search. A cDNA fragment (708 bp) of rat TNF-$\alpha$ was inserted into a phagemid vector, and TNF-$\alpha$ antisense molecules (TNF$\alpha$-LCAS) were produced as single-stranded circular DNA. When introduced into a rat monocyte/macrophage cell line, WRT7/P2, TNF$\alpha$-LCAS was able to ablate LPS-induced TNF-$\alpha$ mRNA to completion. The antisense effect of TNF$\alpha$-LCAS was shown to be sequence-specific because expressions of three control genes ($\beta$-actin, GAPDH and IL-1$\beta$) were not significantly altered by the antisense treatment. Further, TNF$\alpha$-LCAS was found to be highly efficacious as only 0.1 $\mu$g (0.24 nM) of TNF$\alpha$-LCAS was sufficient to block TNF-$\alpha$ expression in 1$\times10^5$ WRT7/P2 cells. I have also observed specific antisense activity in reduction of NF-$\kappa$B gene expression. The results suggest that an antisense sequence as a part of single-stranded circular genomic DNA has a specific antisense activity.

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cDNA Cloning and Developmental Expression of Hemolin in Bombyx mandarina

  • Kang Min Uk;Kim Kyung-A;Lee Jin Sung;Kim Nam Soon;Kang Seok-Woo;Nho Si-kab
    • International Journal of Industrial Entomology and Biomaterials
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    • 제10권2호
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    • pp.101-106
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    • 2005
  • In this study, we describe the Bombyx mandarina hemolin cDNA. A sequence analysis of cDNA revealed a single open reading frame (ORF) of 1233 nucleotides. The deduced 410 amino acid sequence of B. mandarina hemolin contains 4 imunoglobulin (Ig) C-2 type domains. B. mandarina hemolin cDNA showed the highest sequence homology to known those of B. mori. The developmental profile in terms of expression level of hemolin mRNA was determined in the absence of a bacterial challenge. Hemolin mRNA was detected only in mid-gut, but not in hemocytes, fat body, testis, and silkglands. Hemolin mRNA in mid-gut was not detected until the spinning stage of the last instar larva, however, lit dramatically increased at the beginning of spinning and gradually decreased until pupal stage.

cDNA Sequence and mRNA Expression of a Putative Glutathione S-Transferase from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Kim, Jin-Won;Ryu, Kang-Sun;Ahn, Mi-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • 제6권2호
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    • pp.157-162
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    • 2003
  • The glutathione S-transferase (GSTs) are enzymes responsible for the protection of cells from chemical toxicants and oxidative stress. We describe here the cDNA sequence and mRNA expression of a putative GST from the mole cricket, Gryllotalpa orientalis. The G. orientalis GST cDNA sequences comprised of 621 bp encoding 207 amino acid residues. The multiple sequence alignment of G. orientalis GST gene with other known insect GSTs showed several conserved residues that may be essential for the enzymatic activity of the protein. Phylogenetic analysis of the deduced amino acid sequences of G. orientalis GST gene with other insect GST sequences revealed that the G. orientalis GST gene belongs to class I GST, forming a strong monophyletic group (100% bootstrap value) exclusively for class I GSTs from a diverse insect species. Northern blot analysis confirmed midgut-specific expression at transcriptional level, evidencing the midgut as a site for GST synthesis.

구두충(Longicollum pagrosomi)에 감염된 참돔(Pagrus major)의 Heat Shock Protein 70 (Hsp70) 발현 (Expression of a Heat Shock Protein 70 (Hsp70) in Red Seabream Pagrus major Infected with Longicollum pagrosomi)

  • 박형준;민병화
    • 한국수산과학회지
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    • 제51권2호
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    • pp.163-169
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    • 2018
  • This study examined the expression of heat shock protein 70 (Hsp70) in red seabream Pagrus major infected by the, acanthocephalan parasites Longicollum pagrosomi. We cloned the full-length Hsp70 cDNA from the liver of the red seabream. The full-length cDNA had a 1,950 bp open reading frame (ORF) that encoded a protein of 650 amino acids. The deduced amino acid sequence of Hsp70 contained all of the conserved Hsp70 family signature sequences and an adenosine triphosphate (ATP)/guanosine triphosphate (GTP) binding motif, including the EEVD (consensus sequence that terminates in Hsp70 family) consensus sequence. The expression of Hsp70 mRNA was upregulated int the fish head-kidney and liver, as determined by quantitative real-time PCR. We quantified the Hsp70 mRNA expression in normal red seabream and fish infected fish by L. pagrosomi. The expression of Hsp70 mRNA was significantly higher in the infected red seabream. These results suggest that Hsp70 play a role of protection against stress and inflammation caused by the parasite and may help maintain homeostasis.

전기화학법을 이용한 DNA Hybridization 특성 검출 (Detection of DNA Hybridization Characteristics Using Electrochemical methods)

  • 김도균;장정수;권영수
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2002년도 하계학술대회 논문집 C
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    • pp.1569-1571
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    • 2002
  • The determination of DNA hybridization can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, The determination of hybridization is very important for the improvement of DNA detection system. In this study, we report the characterization of the DNA hybridization by the electricalchemical methods. A new electrochemical biosensor is described for voltammetric detection of gene sequence related to probe oligonucleotide of bacterium Escherichia coli O157:H7. The biosensor involves the immobilization of a 18-mer probe oligonucleotide, which is complemetary to a specific gene sequence related to Escherichia coli O157:H7 on a gold electrode through specific adsorption. The probe oligonucleotide was used to determine the amount of target oligonucleotide in solution using mitoxantrone(MTX) as the electrochemical indicators. The cathodic peak currents $(I_{peak})$ of MTX were linearly related to the concentration of the target oligonucleotide sequence in the range $1[{\mu}M]{\sim}0.1[nM]$. The detection limit of this approach was 0.01[nM]. In addition, these indicators were capable of selectivity discriminating against various mismatching condition.

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Identification of Backlash Nonlinear System by use of M-sequence and correlation

  • Kashiwagi, H.;Rong, Li.;Harada, H.
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2000년도 제15차 학술회의논문집
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    • pp.470-470
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    • 2000
  • This paper describes a new method of identifcation of backlash nonlinear systems by use of M-sequence correlation method. In this method, we can obtain not only Volterra kernels of up to 3rd order of the nonlinear system, but also the width of the backlash element from observing the crosscorrelation between the input and the output. Here strictly speaking, a multi-valued nonlinear system such as backlash element can not be expressed by Volterra kernel representation mathematically. But in practice, we encounter many cases where it is difficult to treat them mathematically but they can be controlled from experience. So we here dare to suppose that backlash nonlinear system can be approximated by Volterra kernel representation. Simulations are carried out on a nonlinear system consisting of linear part plus backlash element. And Volterra kernels are measured. The output calculated from the observed Volterra kernels is in good agreement wi th the actual output. And we show that we can obtain the width of backlash element, which is one of the most important parameters, by observing the maximum value of crosscorrelation function between the input M-sequence and the output.

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Molecular Characterization of Porcine DNA Methyltransferase I

  • Lee, Yu-Youn;Kang, Hye-Young;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • 제34권4호
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    • pp.283-288
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    • 2010
  • During normal early embryonic development in mammals, the global pattern of genomic DNA methylation undergoes marked. changes. The level of methylation is high in male and female gametes. Thus, we cloned the cDNA of the porcine DNA methyltransferase 1 (Dnmt1) gene to promote the efficiency of the generation of porcine clones. In this study, porcine Dnmt1 cDNA was sequenced, and Dnmt1 mRNA expression was detected by reverse transcription-polymerase reaction (RT-PCR) in porcine tissues during embryonic development. The porcine Dnmt1 cDNA sequence showed more homology with that of bovine than human, mouse, and rat. The complete sequence of porcine Dnmt1 cDNA was 4,774-bp long and consisted of an open reading frame encoding a protein of 1611 amino acids. The amino acid sequence of porcine DNMT1 showed significant homology with those of bovine (91%), human (88%), rat (76%), and mouse (75%) Dnmt1. The expression of porcine Dnmt1 mRNA was detected during porcine embryogenesis. The mRNA was detected at stages of porcine preimplantation development (1-cell, 2-cell, 4-cell, 8-cell, morula, and blastocyst stages). It was also abundantly expressed in tissues (lung, ovary, kidney and somatic cells). Further investigations are necessary to understand the complex links between methyltransferase 1 and the transcriptional activity in cloned porcine tissues.

N/M/D/F/Fmax 일정계획 문제에서 최적 알고리듬의 개발 (A Development of Optimal Algorithms for N/M/D/F/Fmax Scheduling Problems)

  • 최성운
    • 산업경영시스템학회지
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    • 제13권21호
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    • pp.91-100
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    • 1990
  • This paper is concerned with the development of optimal algorithms for multi-stage flowshop scheduling problems with sequence dependent setup times. In the previous researches the setup time of a job is considered to be able to begin at the earliest opportunity given a particular sequence at the start of operations. In this paper the setup time of a job is considered to be able to begin only at the completion of that job on the previous machine to reflect the effects of the setup time to the performance measure of sequence dependent setup time flowshop scheduling. The results of the study consist of two areas; first, a general integer programming(IP) model is formulated and a nixed integer linear programming(MILP) model is also formulated by introducing a new binary variable. Second a depth-first branch and bound algorithm is developed. To reduce the computational burdens we use the best heuristic schedule developed by Choi(1989) as the first trial. The experiments for developed algorithm are designed for a 4$\times$3$\times$3 factorial design with 360 observations. The experimental factors are PS(ratio of processing time to setup time), M(number of machines), N(number of jobs).

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Detection of Mycobacterium leprae by Real-time PCR Targeting Mycobacterium leprae-Specific Repetitive Element Sequence

  • Jin, Hyun-Woo;Wang, Hye-Young;Kim, Jong-Pill;Cho, Sang-Nae;Lee, Hye-Young
    • 대한의생명과학회지
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    • 제16권2호
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    • pp.127-131
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    • 2010
  • Mycobacterium leprae detection is difficult even with molecular biological techniques due to the low sensitivity of current methodologies. In this report, real-time PCR targeting the M. leprae-specific repetitive element (RLEP) sequence was developed as a new diagnostic tool and evaluated using clinical specimens. For this, M. leprae DNAs were extracted from skin biopsy specimens from 80 patients and analyzed by real-time PCR using TaqMan probe. Then, the detection efficiency of the real-time PCR was compared with that of standard PCR. In brief, the rate of positive detection by the standard PCR and real-time PCR was 32.50% and 66.25%, respectively. The results seemed to clearly show that the TaqMan real-time PCR developed in this study may be a useful tool for sensitive detection of M. leprae from clinical specimens.

한국과 일본 소에 감염된 Theileria 분리주의 small subunit ribosomal 유전자의 동정 및 분석 (Identification and sequence analysis of small subunit ribosomal RNA gene of bovine Theileria isolates from Korea and Japan)

  • 채준석;박진호;권오덕;;;;이주묵
    • 대한수의학회지
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    • 제38권4호
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    • pp.909-917
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    • 1998
  • 한국과 일본의 서로 다른 지역으로부터 소의 Theileria 분리주에 있어서 6가지 type(A부터 E 그리고 H)과 subtype(B1)의 small subunit ribosomal RNA(SSUrRNA) 유전자를 밝혔다. 이들 유전자 염기서열을 비교하여 본 바 염기서열의 위치 212~231, 261~270 그리고 632~690으로 3군데의 hypervariable region이 관찰되었다. SSUrRNA 유전자 염기서열 type A는 한국의 전북 분리주(KCB), 충남 분리주(KCN), 제주 분리주(KCJ)그리고 실험실 보관주(KLS)와 일본의 Shintoku 분리주(JHS)인 5개의 분리주에서 나타났으며, 이 염기서열은 Kenya의 Marula 분리주인 Theileria buffeli의 SSUrRNA 유전자(GenBank accession number Z15106)와 일치하였다. 한국의 경북 분리주(KKB)에서는 type B만이 관찰되었으나 그 외의 분리주에서는 2 type 이상의 유전자 염기서열이 관찰되었다. KCB와 JHS 분리주에서는 type A와 B, 강원 분리주(KKW)에서는 type B와 H, KCN 분리 주에서는 type A, C 및 D 그리고 KCJ 분리주에서는 type A, B, E 및 subtype B1이 관찰되었다. 한국과 일본 소의 Theileria 분리주에 있어서 여러 type의 SSUrRNA 유전자 염기서열이 나타나는 것으로 보아 혼함감염이 되어 있는 것으로 판단된다.

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