• 제목/요약/키워드: insertion sequence

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Genetic Organization of the dhlA Gene Encoding 1,2-Dichloroethane Dechlorinase from Xanthobacter flavus UE15

  • Song, Ji-Sook;Lee, Dong-Hun;Lee, Kyoung;Kim, Chi-Kyung
    • Journal of Microbiology
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    • 제42권3호
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    • pp.188-193
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    • 2004
  • Xanthobacter flavus strain UE15 was isolated in wastewater obtained from the Ulsan industrial complex, Korea. This strain functions as a 1,2-dichloroethane (1,2-DCA) degrader, via a mechanism of hydrolytic dechlorination, under aerobic conditions. The UE15 strain was also capable of dechlorinating other chloroaliphatics such as 2-chloroacetic acid and 2-chloropropionic acid. The dhlA gene encoding 1,2-DCA dechlorinase was cloned from the genomic DNA of the UE15 strain, and its nucleotide sequence was determined to consist of 933 base pairs. The deduced amino acid sequence of the DhlA dechlorinase exhibited 100% homology with the corresponding enzyme from X. autotrophicus GJ10, but only 27 to 29% homology with the corresponding enzymes from Rhodococcus rhodochrous, Pseudomonas pavonaceae, and Mycobacterium sp. strain GP1, which all dechlorinate haloalkane compounds. The UE15 strain has an ORF1 (1,356 bp) downstream from the dhlA gene. The OFR1 shows 99% amino acid sequence homology with the transposase reported from X. autotrophicus GJ10. The transposase gene was not found in the vicinity of the dhlA in the GJ10 strain, but rather beside the dhlB gene coding for haloacid dechlorinase. The dhlA and dhlB genes were confirmed to be located at separate chromosomal loci in the Xanthobacter flavus UE15 strain as well as in X. autotrophicus GJ10. The dhlA and transposase the UE15 strain were found to be parenthesized by a pair of insertion sequences, 181247, which were also found on both sides of the transposase gene in the GJ10 strain. This unique structure of the dhlA gene organization in X. flavus strain UE15 suggested that the dechlorinase gene, dhlA, is transferred with the help of the transposase gene.

Characterization in Terms of the NUX Rule of G-inserted Mutant Hammerhead Ribozymes with High Level of Catalytic Power

  • Kuwabara, Tomoko;Warashina, Masaki;Kato, Yoshio;Kawasaki, Hiroaki;Taira, Kazunari
    • BMB Reports
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    • 제34권1호
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    • pp.51-58
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    • 2001
  • Attempts using in vitro and in vivo selection procedures have been made to search for hammerhead ribozymes that have higher activities than the wild-type ribozyme and also to determine whether other sequences might be possible in the catalytic core of the hammerhead ribozyme. Active sequences selected in the past conformed broadly to the consensus core sequence except at A9, and no sequences were associated with higher activity than that of the hammerhead with the consensus core, an indication that the consensus sequence derived from viruses and virusoids is probably the optimal sequence [Vaish et al. (1997) Biochemistry 36, 6495-6501]. Recently, during construction of ribozyme expression vectors, we isolated a mutant hammerhead ribozyme, with an insertion of G between A9 and G10.1, that appeared to show significant activity [Kawasaki et al. (1996) Nucleic Acids Res. 24, 3010-3016; Kawasaki et al. (1998) Nature 393, 284-289]. We, therefore, characterized kinetic properties of the G-inserted mutant ribozymes in terms of the NUX rule. We demonstrate that the NUX rule is basically applicable to the G-inserted ribozymes and, more importantly, one type of G-inserted ribozyme was very active with $k_{cat}$, value of $6.4\;min^{-1}$ in 50 mM Tris-HCl (pH 8.0) and 10 mM $MgCl_2$ at $37^{\circ}C$.

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Characteristics of the Molecular Epidemiology of CTX-M-Producing Escherichia coli Isolated from a Tertiary Hospital in Daejeon, Korea

  • Kim, Semi;Sung, Ji Youn;Cho, Hye Hyun;Kwon, Kye Chul;Koo, Sun Hoe
    • Journal of Microbiology and Biotechnology
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    • 제26권9호
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    • pp.1643-1649
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    • 2016
  • The aims of this study were to characterize the molecular epidemiological profiles of CTX-M-producing uropathogenic Escherichia coli isolates from a tertiary hospital in Daejeon, Korea, and to investigate the genetic diversity and compare the prevalence of sequence types (STs) in different areas. Extended spectrum β-lactamase-producing E. coli strains isolated from urine were analyzed for CTX-M, integrons, and insertion sequence common regions (ISCRs) by PCR and sequencing. Multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), phylogenetic analysis, and rep-PCR were also used for molecular typing of the isolates. Of 80 CTX-M producers, 31 and 46 expressed CTX-M-15 and CTX-M-14, respectively. MLST analysis indicated that the most prevalent ST was ST131 (n = 34, 42.5%), followed by ST38 (n = 22, 27.5%), ST405 (n = 8, 10.0%), and ST69 (n = 6, 7.5%). Most CTX-M producers harbored class 1 integrons. ST131 strains belonged to phylogenetic group B2 and showed identical rep-PCR patterns, whereas ST69, ST38, and ST405 strains belonged to phylogenetic group D; the ST38 and ST405 strains displayed the same rep-PCR pattern, respectively. ST131 and ST38 isolates showed 21 and 19 distinct types, respectively, by PFGE. In Daejeon, D-ST38 CTX-M-14 producers were relatively more prevalent than in other countries and Korean cities. Our results indicate that CTX-M-producing E. coli isolates belonged mostly to ST131 or ST38 and were more related to hospital-onset than to community-onset infections and that the blaCTX-M gene may vary according to the ST.

Analysis of Delta-V Losses During Lunar Capture Sequence Using Finite Thrust

  • Song, Young-Joo;Park, Sang-Young;Kim, Hae-Dong;Lee, Joo-Hee;Sim, Eun-Sup
    • Journal of Astronomy and Space Sciences
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    • 제28권3호
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    • pp.203-216
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    • 2011
  • To prepare for a future Korean lunar orbiter mission, semi-optimal lunar capture orbits using finite thrust are designed and analyzed. Finite burn delta-V losses during lunar capture sequence are also analyzed by comparing those with values derived with impulsive thrusts in previous research. To design a hypothetical lunar capture sequence, two different intermediate capture orbits having orbital periods of about 12 hours and 3.5 hours are assumed, and final mission operation orbit around the Moon is assumed to be 100 km altitude with 90 degree of inclination. For the performance of the on-board thruster, three different performances (150 N with $I_{sp}$ of 200 seconds, 300 N with $I_{sp}$ of 250 seconds, 450 N with $I_{sp}$ of 300 seconds) are assumed, to provide a broad range of estimates of delta-V losses. As expected, it is found that the finite burn-arc sweeps almost symmetric orbital portions with respect to the perilune vector to minimize the delta-Vs required to achieve the final orbit. In addition, a difference of up to about 2% delta-V can occur during the lunar capture sequences with the use of assumed engine configurations, compared to scenarios with impulsive thrust. However, these delta-V losses will differ for every assumed lunar explorer's on-board thrust capability. Therefore, at the early stage of mission planning, careful consideration must be made while estimating mission budgets, particularly if the preliminary mission studies were assumed using impulsive thrust. The results provided in this paper are expected to lead to further progress in the design field of Korea's lunar orbiter mission, particularly the lunar capture sequences using finite thrust.

Identification of a Gene for Aerobic Growth with a SoxS Binding Sequence in Escherichia coli by Operon Fusion Techniques

  • Lee, Yong-Chan;Kwon, Hyung-Bae;Lee, Sang-Ho;Kwon, Hye-Won;Sung, Ha-Chin;Kim, Joon;Choe, Mu-Hyeon
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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    • pp.1115-1119
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    • 2001
  • Eight Escherichia coli cells with aerobic growth deflects were isolated by the insertion of ${\lambda}placMu53$, a hybrid bacteriophage of ${\lambda}$ and Mu, which created transcriptional fusion to lacZY. Two of these mutants, CLIO and CLl2, were irradiated with UV to obtain specialized transducing phages. The phages that took out the neighboring chromosomal DNA of the related gene responsible for deflective aerobic growth were identified. The in vivo cloned chromosomal sequence revealed that the mutated gene of CLIO was located at min 34.5 on the Escherichia coli linkage map and 1,599,515 on the physical map. The physical map indicated that there were 7 cistrons in the operon. We named this operon oxg10. The promoter sequence of oxg10 exhibited a possible binding site far SoxS, a transcriptional regulator that activates the transcription of various SoxRS regulon genes. Transferring the oxg10:: ${\lambda}placMu53$ mutation into the wild-type strain, RZ4500, resulted in the inhibition of normal aerobic growth, while the salute mutation in strain MO inhibited aerobic cell growth completely. The full operon sequences of oxg10 were cloned from the Excherichia coli genomic library. The mutated gene of CLl2 was identified to be a sucA gene encoding the ${\alpha}$-ketoglutarate dehydrogenase El component in the TCA cycle.

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Intronic Sequences for the Discrimination of Silkworm Strains in Silkworm Powder

  • Kim, Kee-Young;Lee, Eun-Mee;Hong, Mee-Yeon;Kang, Pil-Don;Ryu, Kang-Sun;Kim, Ik-Soo
    • International Journal of Industrial Entomology and Biomaterials
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    • 제15권2호
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    • pp.93-100
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    • 2007
  • After a serial study on the therapeutic efficacy of the powdery silkworm for diabetics was positively resulted in, many powdery silkworm products were on the market in Korea. Up to now, however, no causal method is available to discriminate the strain of silkworms that is a major ingredient for manufacturing powdery silkworm, even though the quality of the powdery silkworm differs greatly by source and origin of strains. We previously were successful in identifying 25 silkworm strains kept in Korea using nine intronic sequences. In this study, we tested the utility of the nine intronic sequences to distinguish the most widely reared silkworm strains originated from Korea and China. Two intron regions, PTTH Intron3 and PTTH Intron3, showed a substantial sequence divergence (mean sequence divergence of 3.13% in PTTH Intron3 and 4.99% in PTTH Intron3). These two intronic sequences provided no identical sequences among the seven strains tested. Thus, these sequences each along can be used to discriminate the seven strains tested in this study. Furthermore, other intron regions, except for VDP Intron4 allowed us to discriminate $2{\sim}4$ strains by strain-specific unique insertion/deletion or substitution.

Identification of DNA Variations Using AFLP and SSR Markers in Soybean Somaclonal Variants

  • Lee, Suk-Ha;Jung, Hyun-Soo;Kyujung Van;Kim, Moon-Young
    • 한국작물학회지
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    • 제49권1호
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    • pp.69-72
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    • 2004
  • Somaclonal variation, defined as phenotypic and genetic variations among regenerated plants from a parental plant, could be caused by changes in chromosome structure, single gene mutation, cytoplasm genetic mutation, insertion of transposable elements, and DNA methylation during plant regeneration. The objective of this study was to evaluate DNA variations among somaclonal variants from the cotyledonary node culture in soybean. A total of 61 soybean somaclones including seven $\textrm{R}_1$ lines and seven $\textrm{R}_2$ lines from Iksannamulkong as well as 27 $\textrm{R}_1$ lines and 20 $\textrm{R}_2$ lines from Jinju 1 were regenerated by organogenesis from the soybean cotyledonary node culture system. Field evaluation revealed no phenotypic difference in major agronomic traits between somaclonal variants and their wild types. AFLP and SSR analyses were performed to detect variations at the DNA level among somaclonal variants of two varieties. Based on AFLP analysis using 36 primer sets, 17 of 892 bands were polymorphic between Iksannamulkong and its somaclonal variants and 11 of 887 bands were polymorphic between Jinju 1 and its somaclonal variants, indicating the presence of DNA sequence change during plant regeneration. Using 36 SSR markers, two polymorphic SSR markers were detected between Iksannamulkong and its somaclonal variants. Sequence comparison amplified with the primers flanking Satt545 showed four additional stretches of ATT repeat in the variant. This suggests that variation at the DNA level between somaclonal variants and their wild types could provide basis for inducing mutation via plant regeneration and broadening crop genetic diversity.

벼 흰잎마름병균(Xanthomonas oryzae pv. oryzae)의 병원성 유전자의 분자유전학적 연구현황 및 비교유전체 분석 (Current Status on Molecular Genetic Study and Comparative Genomic Analysis of Virulence Related Genes in Xanthomonas oryzae pv. oryzae)

  • 강희완;박영진;이병무
    • 미생물학회지
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    • 제44권1호
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    • pp.1-9
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    • 2008
  • 본 논문은 벼 흰 잎마름병균인 Xanthomonas oryzae pv. oryzae(Xoo)의 병원성 유전자의 분자유전학적 연구현황을 기술하고자 한다. 또한 국내 고유 벼 흰 잎마름병균 KACC10331의 유전체해독 정보를 기반으로 다른 Xanthomonas의 유전체와 비교 분석함으로써, Xoo의 주요 병원성 유전자의 분자구조를 구명하고자 한다. 이를 통해 Xoo 고유 병원성 유전자 탐색 및 기능 해석을 위한 기초자료를 제공하는데 목적이 있다. Xoo 유전체에는 5개 과(family)에 속하는 9 type의 Insertion sequence(IS)가 611 copy로 존재하고 있으며, 주로 병원성 관련 유전자군 주위에 많이 분포하고 있는 것으로 나타났다. 현재까지 연구가 수행된 주요 병원성 관련 유전자인 hypersensitive response and pathogenicity (hrp) 유전자, extracellular polysaccharide (EPS) 유전자, extracellular enzyme 유전자, lipopolysaccharides (LPS) 유전자, 그리고 avilulence 유전자의 분자유전학적 연구현황을 기술하였다.

A Yeast MRE3/REC114 Gene is Essential for Normal Cell Growth and Meiotic Recombination

  • Leem, Sun-Hee
    • Journal of Microbiology
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    • 제37권4호
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    • pp.248-255
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    • 1999
  • We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

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키네틱 타이포그래피를 활용한 영화 오프닝타이틀 시퀀스 표현연구(2012 흥행작 중심으로) (Kinetic Typography in Korean Film, 2012 (Study on the movie opening title sequence expression studies using kinetic typography))

  • 방윤경
    • 만화애니메이션 연구
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    • 통권31호
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    • pp.227-248
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    • 2013
  • 컴퓨터의 발달과 함께 영화 오프닝 타이틀 시퀀스도 하루가 다르게 발전하고 있다. 초기에는 타이틀 및 스텝들의 이름을 몇 장의 텍스트로 촬영하여 옵티컬 방식으로 표현하는 방식에서 시작되어 현재는 영화 도입부와 자연스럽게 연결되는 시퀀스부터 영화 중간에 삽입되는 형태, 나아가 2D, 3D 등의 다양한 표현 기법들로 독립적인 형태를 보이고 있다. 이렇게 오프닝타이틀 시퀀스가 하나의 독립적인 예술로 발전되고 있는 것은 짧게는 60초 길게는 10분 남짓의 짧은 영상만으로 영화의 콘셉트를 전달하는 동시에 줄거리의 함축적인 내용을 암시하고 흥미를 유발하는 기능 등 감독의 의지에 따라 다양한 목적으로 영화와의 독립적이면서도, 분리가 아닌 영화와 하나의 유기체 적이며 독창적인 예술의 창조가 가능하기 때문이다. 이 논문에서는 키네틱 타이포그래피의 이론과 분석 방법을 기반으로 2012년도 한국 흥행 영화 10편의 영화 오프닝 타이틀 시퀀스의 제작 방식을 다양한 방법으로 분석하여 앞으로의 나아갈 오프닝 타이틀 시퀀스의 방향을 예측하고 미학적, 기술적인 모델을 제시해 본다.