• Title/Summary/Keyword: terminator

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

  • Ahn, Hong-Il;Shin, Kong-Sik;Woo, Hee-Jong;Lee, Ki-Jong;Kim, Hyo-Sung;Park, Yong-Hwan;Suh, Seok-Cheol;Cho, Hyun-Suk;Kweon, Soon-Jong
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.15-21
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    • 2011
  • We developed 14 transgenic lines of Chinese cabbage (Brassica rapa) harboring the T-DNA border sequences and CryIAc1 transgene of the binary vector 416 using Agrobacterium tumefaciens-mediated DNA transfer. Six lines had single copy cryIAc1 gene and four of them contained no vector backbone DNA. Of the left border (LB) flanking sequences six nucleotides were deleted in transgenic lines 416-2 and 416-3, eleven nucleotides in line 416-9, and 65 nucleotides including the whole LB sequences in line 416-17, respectively. And we defined 499 bp of genomic DNA (gDNA) of transformed Chinese cabbage, and blast results showed 96% homology with Brassica oleracea sequences. PCR with specific primer for the right border (RB) franking sequence revealed 834 bp of PCR product sequence, and it was consisted of 3' end of cryIAc1, nosterminal region and 52 bp of Chinese cabbage genomic DNA near RB. RB sequences were not found and the 58 nucleotides including 21 bp of nos-terminator 3' end were deleted. Also, there were deletion of 10 bp of the known genomic sequences and insertion of 65 bp undefined genomic sequences of Chinese cabbage in the integration site. These results demonstrate that the integration of T-DNA can be accompanied by unusual deletions and insertions both in transgenic and genomic sequences.

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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    • v.3 no.2
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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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Tracking and erosion resistance of polymer for outdoor high voltage insula (초고압 옥외 절연용 고분자 재료의 트래킹 열화특성)

  • Han, Dong-Hee;Park, Hoy-Yul;Kang, Dong-Pil;Kim, In-Sung
    • Proceedings of the KIEE Conference
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    • 1999.07d
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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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    • v.27 no.1
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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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GTD Analysis of Electromagnetic Plane Wave Scattering by Open-Ended Parallel Plate Waveguide with a Slanted Terminator Inside (GTD를 이용한 경사진 벽으로 막힌 평행도파관의 전자파 산란 해석)

  • 선영식;명노훈
    • Journal of the Korean Institute of Telematics and Electronics A
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    • v.29A no.11
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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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    • v.7 no.3
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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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    • v.8 no.6
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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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    • v.4 no.4
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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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Chemical Synthesis of a Human Lysozyme Gene and Expression in Saccharomyces cerervisiae (Human Lysozyme 유전자의 화학적 합성과 Saccharomyces cerevisiae 에서의 발현)

  • 김기운;이승철;황용일
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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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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    • v.1 no.1
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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.