• 제목/요약/키워드: Terminator

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Agrobacterium을 이용하여 형질전환시킨 배추에서 T-DNA Right Border 인접염기서열 분석 (Analysis of right border flanking sequence in transgenic chinese cabbage harboring integrated T-DNA)

  • 안홍일;신공식;우희종;이기종;김효성;박용환;서석철;조현석;권순종
    • Journal of Plant Biotechnology
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    • 제38권1호
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    • pp.15-21
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    • 2011
  • Agrobacterium tumefaciens와 운반체 416을 이용하여 cryIAc1 유전자가 형질전환된 배추 14개체를 선발하였다. Southern blot을 통하여 분석한 결과 1 사본 유전자가 도입된 개체가 6개이었으며 backbone 염기서열이 포함되지 않은 경우는 4개체이었다. LB 인접서열 분석 결과, 416-2과 416-3은 23 bp의 LB 부위가 남아있는 동일한 염기서열을 보였고, 416-9 경우 15 bp가 남았으며, 416-17 경우 LB를 포함하여 안쪽의 염기서열의 최대 36 bp의 결실이 확인되었다. T-DNA의 염기서열을 제외한 배추 gDNA의 499 bp의 LB 인접염기서열은 B. oleracea의 염기서열과 96% 상동성을 가진 유전자로 확인되었다. RB border 인접서열 분석용 primer를 제작하여 PCR을 수행한 결과 모두 834 bp의 염기서열을 확인하였고, vector의 구성 요소 중 cryIAc1의 3' 말단 부위, nos-terminator 부위와 52 bp 및 배추 gDNA RB 인접염기서열로 확인되었다. RB 염기서열은 확인할 수 없었으며 nos-terminator 3' 말단의 21개의 염기를 포함하여 모두 58개의 염기서열이 결실된 것을 확인하였다. 형질전환식물체를 제작할 경우 발생하는 전이유전자나 식물의 염색체에 염기 결실은 매우 다양한 형태로 나타난다. 이번 실험의 경우, 전이유전자가 삽입된 위치에서 약 10 bp의 배추의 gDNA 염기가 결실된 것이나 삽입된 전이유전자의 RB 말단부분이 결실된 것은 예상할 수 있는 결과였지만, 특히 전이유전자의 말단인 nos-terminator 3' 끝에 65 bp 정도의 배추의 다른 유전자가 삽입되어 있다는 것을 확인하였고 이는 기대하지 않았던 결과였다. 삽입된 다른 배추유전자의 절편은 앞으로 더 많은 연구를 통하여 위치와 기능확인이 필요할 것이다.

Nucleotide Sequence of a Bacteriolytic Enzyme Gene from Alkalophilic Bacillus sp.

  • Jung, Myeong-Ho;Ohk, Seung-Ho;Yum, Do-Young;Kong, In-Soo;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제3권2호
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    • pp.73-77
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    • 1993
  • The nucleotide sequence of Bacillus sp. bacteriolytic enzyme gene, lytP and its flanking regions were determined. A unique open reading frame for a protein of Mw. 27, 000, and a putative terminator sequence, were found behind a concensus ribosome binding site located 8 nt upstream from ATG start codon. The primary amino acid sequence deduced from nucleotide sequence revealed a putative protein of 255 amino acid residues with an Mw. of 27, 420. No significant homology could be found between the amino acid sequence of Bacillus sp. bacteriolytic enzyme and that of other cell wall hydrolases.

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초고압 옥외 절연용 고분자 재료의 트래킹 열화특성 (Tracking and erosion resistance of polymer for outdoor high voltage insula)

  • 한동희;박효열;강동필;김인성
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 1999년도 하계학술대회 논문집 D
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    • pp.1578-1580
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    • 1999
  • Silicone rubber is being used for the housing material of outdoor high voltage insulators such as composite insulator, bushing, surge arrestor and cable terminator because of good tracking and erosion resistance, good hydrophobicity and recovery of hydrophobicity, and chemical stability. In this paper, tracking and erosion resistance of silicone rubber having fluids and different ATH contents were examined. Fluids were selected under the consideration of their molecular weight and chemical structure, expecting the high migration rate, the good pollutant encapsulation, and the long period with good hydrophobicity. Good tracking and erosion resistance and arc resistance have been achieved for the silicone rubber above ATH content 130 phr.

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Voltage-Mode 1.5 Gbps Interface Circuits for Chip-to-Chip Communication

  • Lee, Kwang-Jin;Kim, Tae-Hyoung;Cho, Uk-Rae;Byun, Hyun-Geun;Kim, Su-Ki
    • ETRI Journal
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    • 제27권1호
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    • pp.81-88
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    • 2005
  • In this paper, interface circuits that are suitable for point-to-point interconnection with an over 1 Gbps data rate per pin are proposed. To achieve a successful data transfer rate of multi-gigabits per-second between two chips with a point-to-point interconnection, the input receiver uses an on-chip parallel terminator of the pass gate style, while the output driver uses the pullup and pulldown transistors of the diode-connected style. In addition, the novel dynamic voltage level converter (DVLC) has solved such problems as the access time increase and valid data window reduction. These schemes were adopted on a 64 Mb DDR SRAM with a 1.5 Gbps data rate per pin and fabricated using a 0.10 ${\mu}m$ dual gate oxide CMOS technology.

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GTD를 이용한 경사진 벽으로 막힌 평행도파관의 전자파 산란 해석 (GTD Analysis of Electromagnetic Plane Wave Scattering by Open-Ended Parallel Plate Waveguide with a Slanted Terminator Inside)

  • 선영식;명노훈
    • 전자공학회논문지A
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    • 제29A권11호
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    • pp.19-24
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    • 1992
  • In this paper, a high frequency method is developed which combines the uniform Geometrical Theory of Diffraction(GTD) and the Aperture Integration(AI) to analyze electromagnetic plane wave scattering by a perfectly-conducting, open-ended, semi-infinite parallel plate waveguide with a uniform layer of absorbing material on its inner wall, and with a slanted planar termination inside. In this method, first, the field of an arbitary point inside the paraller plate waveguide is computed by the GTD. Second, the field scattered into exterior region by the waveguide is found using the equivalent current, which can be obtaind from the aperture field of the waveguide and using the AI. Numerical results based on this GTD method are presented and compared with those based on the mode matching method.

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Random Insertional Mutagenesis with Subtracted cDNA Fragments in Arabidopsis thaliana

  • Euna Cho;Kwon, Young-Myung;Lee, Ilha
    • Journal of Photoscience
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    • 제7권3호
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    • pp.103-108
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    • 2000
  • We have evaluated a new mutagenesis strategy called random insertional mutagenesis with subtracted cDNA fragments. The cDNAs from long day Arabidopsis plants were subtracted by cDNAs from short day plants using PCR based cDNA subtraction. The subtracted cDNAs were inserted between 35S promoter and 3'-NOS terminator regardless of orientation. When the cDNA library was used for the random insertion into Arabidopsis genome by Agrobacterium-mediated transformation, approximately 15% of transformants showed abnormal development in leaf, floral organ, shoot apex. When 20 mutants were analyzed, 12 mutants showed single cDNA fragment insertion and 8 mutants showed more than 2 transgene insertions. Only two mutants among 12 mutants that have single cDNA insert showed consistent phenotype at T2 generation, suggesting the genetic instability of the mutants.

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Construction of a Secretory Expression Vector Producing an $\alpha$-Amylase of Yeast, Schwanniomyces occidentalis in Saccharomyces

  • Shin, Dong-Jun;Park, Jong-Chun;Lee, Hwanghee-Blaise;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • 제8권6호
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    • pp.625-630
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    • 1998
  • Using a modified yeast secretory expression vector, $\alpha$-amylase of Schwanniomyces occidentalis was produced from Saccharomyces cerevisiae. The expression vector contains the a-amylase gene (AMY) harboring its own promoter without the regulatory region and the adenine base at the -3 position from the ATG start codon, its own signal sequence, CYC1 transcription terminator, and SV40 enhancer. The expressed $\alpha$-amylase activity from cells carrying the plasmid was approximately 26 times higher than that from the cells harboring an unmodified plasmid. When Saccharomyces diastaticus was transformed with this modified vector, a 2.5 times higher level of amylolytic activity than that from Sch. occidentalis was observed.

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Construction of Recombinant Xanthomonas campestris Strain Producing Insecticidal Protein of Bacillus thuringiensis

  • Shin, Byung-Sik;Koo, Bon-Tag;Choi, Soo-Keun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제4권4호
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    • pp.285-289
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    • 1994
  • An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated into the chromosome of a xanthan-producing bacterium, Xanthomonas campestris XP92. The cryIA(c) gene expression cassette was constructed that placed the gene between the trc promoter and rrnB transcriptional terminator. The $lacl^q$ gene was also included to prevent the expression of cryIA(c) gene in X campestris cells. Southem blot analysis confirmed the integration of the cryIA(c) gene expression cassette in chromosome of X campestris XP92 transconjugant. Expression of the insecticidal crystal protein was confirmed by Western blot analysis and bioassay against the larvae of Hyphantria cunea (Lepidoptera: Arctiidae) and Plutella xylostella (Lepidoptera:Plutellidae).

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Human Lysozyme 유전자의 화학적 합성과 Saccharomyces cerevisiae 에서의 발현 (Chemical Synthesis of a Human Lysozyme Gene and Expression in Saccharomyces cerervisiae)

  • 김기운;이승철;황용일
    • 한국미생물·생명공학회지
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    • 제23권2호
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    • pp.138-144
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    • 1995
  • The cDNA, encoding human lysozyme (HLY) which was isolated from a human placenta cDNA library, has been well characterized (Yoshimura et al., 1988). Based on the communication, we have prepared an artificial HLY gene from chemically synthesized 38-oligomer with high codon usage in Saccharomyces cerevisiae. For directing the synthesis and secretion of HLY in S. cerevisiae, an expression vector, pHKl was constructed by inserting the HLY gene, containing a synthetic HLY secretion signal sequence, between the yeast GAP promoter and PH05 terminator. From a lysoplate assay, we have confirmed an yeast transformant harboring a pHK1 which makes a clearing zone on the overlayed Micrococcus luteus. This result means a chemically synthesized HLY gene which was normally expressed and secreted in yeast.

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A Small Cryptic Plasmid pZMO1 of Zymomonas mobilis ATCC10988

  • Kang, Hyung-Lyun;Kang, Hyen-Sam
    • Genomics & Informatics
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    • 제1권1호
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    • pp.55-60
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    • 2003
  • The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed $69\%$ identity with Shigella sonnei plasmid, pKYM and $61\%$ identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed $75\%$ identity and $90\%$ similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.