• Title/Summary/Keyword: Terminator

Search Result 122, Processing Time 0.024 seconds

Searching Method for New Small RNA in Bacillus subtilis Using Bioinformation (생물정보를 이용하여 바실러스 서브틸리스에서 새로운 Small RNA를 예측하는 방법)

  • Lee, Sang-Soo
    • The Journal of Natural Sciences
    • /
    • v.18 no.1
    • /
    • pp.47-53
    • /
    • 2007
  • In order to find novel sRNA in Bacillus subtilis which would be used to adapt to several conditions, we searched the whole genome of Bacillus subtilis using the following procedure. At first, the locations of recognition sequence of transcription factors such as PerR, OhrR, Fur and Zur were searched in the intergenic region of Bacillus subtilis genome and the locations of rho independent transcription terminator sites were also determined. Based on the information of these locations, the sRNA candidates were chosen by close locations (less than 300 bp) between the recognition site of transcription factors and rho independent transcription terminator site. Than transcription promoter sites were searched in the region of previously identified sRNA candidates and 5 PerR, 1 OhrR, 1 Fur and 1 Zur regulated good sRNA candidates were found.

  • PDF

Optimization of Environmental Conditions for Hirudin Production from Recombinant Saccharomyces cerevisiae (재조합 효모를 이용한 Hirudin 발효생산조건의 최적화)

  • 이동훈;서진호
    • KSBB Journal
    • /
    • v.9 no.1
    • /
    • pp.8-15
    • /
    • 1994
  • The research has been carried out to optimize a recombinant S. cerevisine fermentation process for the production of an anticoagulant hirudin. The structural gene coding for hirudin was combined with the GAL10 promoter for controlled expression, the MFal signal sequence for hirudin secretion, and the GAL7 terminator for transcriptional termination. Growth medium composition and environmental conditions were optimized for maximizing cell growth and final hirudin concentration. The optimized conditions included yeast extract 40g/$\ell$, casamino acid 5g/$\ell$, g1ucose 20g/$\ell$, galactose 30g/$\ell$, DO 50% and temperature $30^{\circ}C$. These conditions yielded the specific cell growth rate of $0.13hr^{-1}$, the final cell density of 30g cell/$\ell$ and the final hirudin concentration of 64mg/$\ell$ in the batch fermentation with a 2.5$\ell$ jar fermentor.

  • PDF

Detection of Transgenic Rice Containing CrylAc Gene Derived from Bacillus thuringiensis by PCR

  • Kim, Jae-Hwan;Jee, Sang-Mi;Park, Cheon-Seok;Kim, Hae-Yeong
    • Food Science and Biotechnology
    • /
    • v.15 no.4
    • /
    • pp.625-630
    • /
    • 2006
  • Polymerase chain reaction (PCR) method was developed for the specific detection of insect-resistant rice containing cry1Ac gene derived from Bacillus thuringiensis (Bt). Primers were designed from the 35S promoter, NOS terminator, cry1Ac gene, and sucrose phosphate synthase (SPS) for general screening of Bt rice. By sequencing the PCR products from the two putative kinds of Bt rice, we designed a specific primer from the junction region between the cry1Ac gene and the NOS terminator that had been inserted into Bt rice. The construct-specific primer was employed to amplify a 147 bp product in the two lines of Bt rice. No amplified products were observed from the other Bt crops with various Bt genes introduced. In qualitative PCR analysis, the limit of detection was 0.005 ng from genomic DNA of Bt rice. In addition, PCR analysis was performed on 64 kinds of rice presently available in the Korean market, and no Bt rice was detected. This method presented in this paper can be used as a highly sensitive and specific detection method of Bt rice.

Primer for the Potato Specific Internal Control DNA and Screening Method for the Genetically Modified Potatoes by Competitive Duplex-PCR (감자 특이 Internal Control DNA 증폭용 Primer와 이를 이용한 유전자 변형 감자의 경쟁적 이중 PCR 검정법)

  • Seo, Hyo-Won;Yi, Jung-Yoon;Cho, Hyun-Mook;Kim, Sung-Yeul
    • Journal of Plant Biotechnology
    • /
    • v.29 no.4
    • /
    • pp.235-240
    • /
    • 2002
  • We report the new method for the screening of genetically modified potato by competitive duplex-PCR using the potato specific single oligomer primer for the internal control and CaMV 35S promoter or NOS terminator specific primers. The single oligomer primer (rAGU4A) amplify the potato specific internal control band from the homozygous potato genomic DNA in the RAPD profiles of all analyzed potato varieties. The 530 bp internal control DNA was amplified independently to CaMV 35S promoter or NOS terminator DNA and identified as repetitive or microsatellite DNA of potato (AF541972). With this new technique, the transgenic potatoes which were transformed with vectors contained the different foreign genes are analyzed. In case of the commercialized transgenic potato varieties, 'Hew Leafs', those two genetic factors are used for promoter and terminator respectively So, this new PCR technique should be a promising method of cost effective and accurate screening for the commercialized GM potatoes on market.

Introduction of Shiva Gene into tobacco and Potato Using Tissue-Specific Tomato PAL Promoter (조직특이성 promoter를 이용한 Shiva 유전자의 식물체내 도입)

  • 이정윤;이신우;박권우
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.2
    • /
    • pp.109-113
    • /
    • 1998
  • In this study we tried to transform an antimicrobial peptide gene (Shiva) under the promoter of tomato phenylalanine ammonia-lyase (tPAL5) into tobacco and potato plants. Antimicrobial peptide gene was isolated originally from giant silk moth (Hyalophora cecropia) and modified ie nucleotide sequence to increase antimicrobial activity. Transgenic tobacco plants were regenerated and their seeds were tested on the media containing kanamycin (500 mg/L). The results of PCR amplification and genomic Southern blot hybridization confirmed the integration of construct (tPAL5 promoter-Shiva-NOS-GUS-NOS) into chromosome. We observed that one of the transgenic tobacco plants showed chromosome rearrangement when integrated. In case of potato transformation, the efficiency of regeneration was maximized at the medium containing Zeatin 2mg/L, NAA 0.01mg/L, GA$_3$ 0.1mg/L. We also observed the high expression of GUS (${\beta}$-glucuronidase) enzyme which was located next to the terminator sequence of nopaline synthase gene (NOS) in the vascular tissue of stem, leaves of transgenic potatoes. This result suggested that a short sequence of Shiva gene (120 bp) and NOS terminator sequence might be served as a leader sequence of transcript when translated.

  • PDF

ETRI신기술-다기능 ISDN PC 카드 기술

  • Electronics and Telecommunications Research Institute
    • Electronics and Telecommunications Trends
    • /
    • v.14 no.4 s.58
    • /
    • pp.129-130
    • /
    • 1999
  • N-ISDN의 대중화 및 활성화를 위해서 연구 개발한 회선 및 패킷 통신 기능을 지원하는 N-ISDN 접속용 PC 카드로서, NT(Network Terminator)및 TA(Terminal Adapter) 기능을 하며, PC에서 다중 통신이 가능하도록 한다. 음성 통화중 다양한 ISDN 서비스를 이용 가능하도록 하며, 64kbps로 인터넷 접속을 할 수 있다. 또한 전용선 없이 ISDN을 통해 64kbps로 패킷망에 접속하여 데이터 통신이 가능하도록 하는 등 협대역 ISDN 통신 기능을 지원한다.

  • PDF

Development of a Plant Transformation Vector, pKCHI (고등식물 형질전환용 유전자 운반체 pKCHI의 개발)

  • 정상호
    • Journal of Plant Biology
    • /
    • v.32 no.1
    • /
    • pp.23-32
    • /
    • 1989
  • We have developed a plasmid vector, pKCH1, for the purpose of higher plant transformation. It contains the promoter region of cauliflower mosaic virus 35S transcript (P35s) and the terminator region of nopaline synthase gene (Tnos) with unique cloning sites, Bam HI and Xba I, between them. After inserting a foreing gene at the cloning sites, P35s-foreign gene-Tnos cassette can be recovered by using a restriction enzyme Hind III.

  • PDF

A Uniform GTD and Aperture Integration Analysis of the Electromagnetic Scattering by a Semi-infinite Parallel Plate Waveguide with an Interior Termination and Lossy Inner Walls (Uniform GTD와 Aperture Integration을 이용한 내부에 Terminator가 있는 평면도파관의 전자기파의 산란)

  • Myung, N.H.
    • Proceedings of the KIEE Conference
    • /
    • 1987.07a
    • /
    • pp.105-109
    • /
    • 1987
  • A solution which combines ray and aperture integration(AI) techniques is presented for the problem of electromagnetic plane wave scattering by an open-ended, perfectly-conducting, semi-infinite parallel plate waveguide with a thin, uniform layer of lossy or absorbing material on its inner walls, and with a simple planar termination inside. Numerical results are given for the fields outside the waveguide.

  • PDF

Optimization of SNP Genotyping Assay with Fluorescence Polarization Detection

  • Cai Chun Mei;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.50 no.5
    • /
    • pp.361-367
    • /
    • 2005
  • Single nucleotide polymorphisms (SNPs) are valuable DNA markers due to their abundance and potential for use in automated high-throughput genotyping. Numerous SNP genotyping assays have been developed. In this report, one of effective and high throughput SNP genotyping assays, which was named the template-directed dye-terminator incorporation with fluorescence polarization detection (FP-TDI) was described. Although the most of this assay succeed, the objective of this work was to deter­mine the reasons for the failures, find ways to improve the assay and reduce the running cost. Ninety $F_2$-derived soybean, Glycine max (L.) Merr., RILs from a cross between 'Pureunkong' and 'Jinpumkong 2' were genotyped at four SNPs. FP measurement was done on $Victot^3$ microplate reader (perkinelmer Inc., Boston, MA, USA). Increasing the number of thermal cycles in the single-base extension step increased the separation of the FP values between the products corresponding to different genotypes. But in some assays, excess of heterozygous genotypes was observed with increase of PCR cycles. We discovered that the excess heterozygous was due to misincorporation of one of the dye­terminators during the primer extension reaction. After pyrophosphatase incubation and thermal cycle control, misincoporation can be effectively prevented. Using long amplicons instead of short amplicons for SNP genotyping and decreasing the amount of dye terminator and Acyclopol Taq polymerase to 1/2 or 1/3 decreased the cost of the assay. With these minor adjustments, the FP-TDI assay can be used more accurately and cost-effectively.