• Title/Summary/Keyword: ORF 분석

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Diagnosis of the ORF Virus Using a Mixture of Sieving Gel Matrixes in Microchip Gel Electrophoresis (마이크로칩젤 전기영동에서 충진젤 혼합물을 이용한 ORF 바이러스의 진단)

  • Kim, Yun-Jeong;Chae, Joon-Seok;Kang, Seong-Ho
    • Journal of the Korean Chemical Society
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    • v.48 no.5
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    • pp.483-490
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    • 2004
  • We have developed a novel polymerase chain reaction (PCR)-microchip gel electrophoresis (MGE) method based on the sieving gel mixture of commercially available poly(vinylpyrrolidone) (PVP) and hydroxy ethyl cellulose (HEC) for the rapid detection and diagnosis of the orf virus (ORFV) from Korean indigenous goat. After amplification of 594-bp DNA fragment from the B2L gene of ORF virus, the amplicon was analyzed by the MGE separation. The glass microfluidic chip (64 mm total length (36 mm effective length)${\times}$90 ${\mu}$m width${\times}$20 ${\mu}$m depth) allowed the fast detection and diagnosis of ORFV in the mixture of 1.0% PVP ($M_r$ 360,000) and 1.0% HEC ($M_r$250,000) as a sieving matrix with better resolution and reproducibility of DNA fragments. Under the electric field of 277.8 V/cm, the 594-bp DNA was analyzed within 4 min. Compared to traditional slab gel electrophoresis, the PCR-MGE method was twenty times faster and an effective clinical method for the quantitative analysis of ORFV.

Isolation and sequence analysis of a small cryptic plasmid from Lactobacillus farciminis KCTC3681 (Lactobacillus farciminis로부터 미지의 작은 플라스미드의 분리와 염기서열 분석)

  • Lee, Eun-Mo;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.28 no.B
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    • pp.53-57
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    • 2008
  • From the extensive screening for small cryptic plasmid among about 23 lactic acid bacteria (LAB), 2.4 kb of cryptic plasmid was isolated from Lactobacillus farciminis strain KCTC 3681 and named as pLF24. The plasmid pLF24 was a circular molecule of 2,396 base-pairs in length with a G+C content of 38%. Two protein-coding sequences could be predicted. ORF1 and ORF2 showed homologies to plasmids of gram-positive bacteria. The replication protein coded by ORF2 and the plus origin, were similar to replication regions of other gram-positive bacteria as shown in plasmids such as pLH2, pLS141-1 and pLC2. The nucleotide sequence of pLF24 was deposited into Genbank data base with an accession number of EU429343. The newly isolated plasmid can be used for construction of shuttle vector in Lactobacillus bacteria.

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Complete Genome Analysis of Spodoptera exigua Nucleopolyhedrovirus Isolated in Korea (한국에서 분리된 파밤나방 핵다각체병 바이러스의 전체 유전체 분석)

  • Jae Bang, Choi;Hyun-Soo, Kim;Soo-Dong, Woo
    • Korean journal of applied entomology
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    • v.61 no.3
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    • pp.449-460
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    • 2022
  • The morphology and whole genome sequence of Spodoptera exigua nucleopolyhedrovirus K1 (SeNPV-K1) isolated in Korea were analyzed for the use as an eco-friendly control source against S. exigua. The polyhedra of SeNPV-K1 was amorphous with a size of 0.6-1.8 ㎛, and there was no external difference with the previously reported SeNPV. As a result of analyzing the nucleotide sequence of the whole genome, it was composed of 135,756 bp, which is 145 bp more than that of the previously reported SeNPV. The G+CG+C content was 44% and there were 6 homologous repeated sequences, so there was no significant difference from the previous report. As a result of ORF analysis, SeNPV-K1 had 137, two fewer than those previously reported, and 4 ORFs present only in SeNPV-K1 were confirmed. These 4 ORFs are non-essential genes and were not considered to have a significant influence on the characteristics of the SeNPV. The genome vista analysis showed that the overall sequence similarity between SeNPV-K1 and the previously reported SeNPV was very high. The whole genome of SeNPV-K1 analyzed for the first time in Korea was found to be similar to the previously reported SeNPV, but it was confirmed that it was a novel resource in Korea with different isolate.

Functional Analysis of BCTV ORF L4 by Site-directed Mutagenesis (Site-directed mutagenesis를 이용한 BCTV ORF L4의 기능 분석)

  • 박을용;이석찬
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.513-518
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    • 1998
  • Beet curly top virus (BCTV) mutant has been constructed in vitro that contain G-to-T transversions at nucleotide 2727 within overlapping open reading frames (ORFs) L1 and L4. The mutations introduce termination codon in ORF L4 without affecting the amino acid encoded by ORF L1. When agroinoculated into Arabidopsis thaliana the mutant caused mild stunting and stem curling, but not the callus induction and hyperlasia on infected tissues of Sei-O ecotype. However, this mutant was not infectious on Col-O. Levels of single stranded DNA forms were similar in mutant and wild type BCTV infections. The DNA quantitation data showed that the DNA of BCTV-L4 mutant virus was accumulated in shoot tips, infection origin and roots with similar levels to those of wild type virus infected. Three tissues of asymptomatic ecotype Col-O also had as much as virus DNA from wild type virus infections. In both ecotypes infected with BCTV-Logan and BCTV-L4 mutant, root tissues contained more virus DNA than any other tissues by the Southern hybridization data. The results suggest that ORF L4 encodes a functional protein that is a major determinant of pathogenesis that might affect the hyperplastic response of the host to BCTV infection.

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Unification System for Analysis of DNA Sequence (DNA 서열 분석을 위한 통합 시스템)

  • Song, Young-Ohk;Chang, Duk-Jin
    • The Journal of the Korea Contents Association
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    • v.11 no.3
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    • pp.65-72
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    • 2011
  • We stand at real world that some practical use method of gene information appears in succession by entrance on the stage of advanced techonlogy. As a lot of studies and development are achieved based on analysis of bio data, necessity of a tool that can help correct interpretation of data is required more and more in a lot of targets of bioinformatics to search new relation and information are established. In this paper, we are offered in existing I wish to offer user a more convenient study tool developing system that can supplement shortcomings of various tools for data analysis. So we've designed to offer in united environment that is not environment that is parted ORF driving out, bio information retrieval and work of similarity comparison lamp to work for bio data analysis and offers lacking consecutiveness in existing analysis system.

Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

Cloning, Sequencing and Comparison of Genes for early Enzymes of the Protocatechuate (ortho-Cleavage) Pathway in Pseudomonas putida (Pseudomonas putida의 Protocatechuate 경로에 관여하는 초기 효소들의 유전자의 클로닝 및 염기서열 분석비교)

  • Hong, Bum-Shik;Shin, Dong-Hoon;Kim, Jae-Ho
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.472-476
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    • 1996
  • The major portions of two DNA fragments, one from degradative plasmid, pRA4000 from Pseudomonas putida NCIMB 9866, and the other from degradative plasmid, pRA500 from P. putida NCIMB 9869, which harbor the structural genes for the flavoprotein (pchF) and cytochrome (pchC) subunits of p-cresol methylhydroxylase (PCMH), have been sequenced. The DNA and deduced amino acid sequences for pchC and pchF have been published. In these fragments, a coding region (dhal) for an aldehyde dehydrogenase has been identified. It is proposed that this gene encodes for the aldehyde dehydrogenase which converts p-hydroxybenzyaldehyde to p-hydroxybenzoate. p-Hydroxybezealdehyde is the product of oxidation of p-cresol by PCMH. The fragment from P. putida 9869 also harbors the genes for the ${\alpha}$ (pcaG) and ${\beta}$ (pcaH) subunits of protocatechuate 3,4-dioxigenase. The fragment from 9866 does not have any portion of these genes in the corresponding region A possible open reading frame (ORF) between pchC and pchF is seen for both clones, and a second putative open reading frame (ORF') also exists in the 9866 clone. The gene organizations are dhal-pchC-ORF-pchF-pcaGH for the DNA fragment from 9869, and ORF-dhal-pchC-ORF-pchF for the DNA fragment from 9866.

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Characterization of the Repetitive Sequences Present in the ORF25 Genomic Region of Megalocytiviruses from Ornamental Fishes (관상어로부터 분리한 Megalocytiviruses에서 나타나는 ORF25 유전자 부위의 반복서열 특성 분석)

  • Jin, Ji-Woong;Nam, Jeong-Hee;Kim, Kwang-Il;Hong, Su-Hee;Byon, Ju-Young;Jeong, Hyun-Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.4
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    • pp.352-358
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    • 2011
  • The presence of ISKNV-like viruses in various freshwater ornamental fish species imported from Asia was confirmed by polymerase chain reaction(PCR) amplification of the ATPase(adenosine triphosphatase) gene. Interestingly, molecular analyses of the Open Reading Frame 25(ORF25) region of these isolates based on the ISKNV(Infectious spleen and kidney necrosis virus) genome revealed the presence of various repetitive sequences. ORF25 repeat sequence length had no effect on cumulative mortality of rock bream Oplegnathus fasciatus challenged with tissue homogenates of infected pearl gourami, Trichogaster leeri; silver gourami, Trichogaster microlepis; blue gourami, or Trichogaster trichopterus. All isolates induce cumulative mortalities after 12 days of infection, confirming that ORF25 polymorphism did not affect the pathogenicity of ornamental fish megalocytiviruses that cross infect rock bream, a seawater fish. Also, no statistically significant differences in spleen index or viral copy number in infected tissues was detected between isolates with varying ORF25 repeat sequence lengths. However, further studies are necessary to fully characterize the functional characteristics of these polymorphisms in megalocytivirus disease in ornamental fishes.

Analysis of a Putative DNA Polymerase I gene in Brevibacterium ammoniagenes. (Brevibacterium ammoniagenes의 DNA Polymerase I 유사 유전자의 분석)

  • 오영필;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.105-110
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    • 2002
  • The sequence of 3,221 nucleotides immediately adjacent to rpsA gene encoding 30S ribosomal protein S1 of Brevibacterium ammoniagenes was determined. A putative open reading frame (ORF) of 2,670 nucleotides for a polypeptide of 889 amino acid residues and a TAG stop codon was found, which is located at a distance of 723 nucleotides upstream from rpsA gene with same translational direction. The deduced amino acid sequence of the ORF was found to be highly homologous to the DNA polymerase I of Streptomyces griseus (75.48%), Rhodococcus sp. ATCC 15963 (56.69%), Mycobacterium tuberculosis (55.46%) and Mycobacterium leprae (53.99%). It was suggested that the predicted product of the ORF is a DNA polymerase I with three functional domains. Two domains of 5 → 3 exonuclease and DNA polymerase are highly conserved with other DNA polymerase I, but 3 → 5 exonuclease domain is less conserved.

Design and Implementation of an Intelligent Multiple DNA Sequence Translation Tool (지능적 다중염기서열 변환 도구의 설계 및 구현)

  • Lee Hye-Ri;Lee Geon-Myeong;Lee Chan-Hui;Lee Seong-Deok;Kim Seong-Su
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2006.05a
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    • pp.37-40
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    • 2006
  • 계통분석을 하는 생물학자들은 관련된 분석대상에 대한 정보를 확보하여 비교분석하기 위해 NCBI 등으로부터 염기서열을 확보하여 아미노산 서열로 변환하는 작업을 수행하게 된다. 많은 서열 데이터에 대해서 데이터베이스로부터 데이터를 검색하고 이를 변환하는 작업을 순차적으로 분석자가 관여하여 작업하는 것이 현재 분석환경이다. 따라서 본 논문에서는 분석의 효율성을 향상시키기 위해, 관심서열의 등록번호(Accession Number) 리스트를 입력하면 해당 서열에 대항 정보를 NCBI로부터 웹로봇을 통해 자동으로 확보한 다음, 확보된 염기서열 전체를 아미노산 서열로 자동 변환하여 가장 긴 ORF(Open Reading Frame)을 추천해주기 위해 설계된 지능형 다중 염기서열 변환 도구에 대해서 소개한다.

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