• Title/Summary/Keyword: 제한 효소 지도 작성

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Molecular Cloning of $\beta$-Galactosidase Gene from Neisseria lactamica 2118 into Escherichia coli MC 1061 (Neisseria lactamica 2118의 $\beta$-galactosidase 유전자의 대장균으로의 클로닝)

  • Lee, Jong-Su
    • The Journal of Natural Sciences
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    • v.5 no.1
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    • pp.37-45
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    • 1992
  • The gene coding for $\beta$-galactosidase of Neisseria lactamica 2118 was cloned into Escherichia coli MC 1061. The isolated 6.5 Kb EcoR I fragement and 7.2 Kb BamH I fragment of chromosomal DNA in Southern hybridization were ligated to a vector plasmid pBR322 and then transformed into Escherichia coli MC 1061 cells. Finally, I obtained three clones as $\beta$-galactosidase positive clone by colony hybridization and Southern hybridization($\beta$-galactosidase probe: lac Z gene of pMC1871). Three recombinant plasmids(pNL.13. 17 and 24) were found to contain the 7.2Kb BamH I fragment originated from Neisseria lactamica 2118 chromosomal DNA by Southern hybridization and pNL 24 was showed high homology to probe especially and also its physical map was constructed.

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Genetic and molecular analysis of the R-mb gene from maize (옥수수 R-mb 유전자의 유전분석과 그의 구조)

  • 윤필용;유삼규;송원용;윤충효;임용표
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.161-165
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    • 1997
  • The R-mb locus of maize is one of several genes that encode tissue-specific transcriptional regulator for the anthocyanin biosynthesis in plant parts and the aleurone layer in seeds. We found that the seed pigment frequencies gradually decreased at selfed progenies of the R-mb genetic stocks. In order to analyze the genomic structure of R-mb locus components, genomic Southern blot was performed by using R specific probe, pR-nj:1. Two bands were detected at the size of about 3.9 and 7.75kb. Five R-mb positive clones (mb-II, III, V,Ⅵ, and Ⅶ) were obtained by screening of maize genomic λFIXII library using R specific probe pR-nj:1. We constructed the restriction map of clone mb-II (7.75Kb positive) and mb-Ⅵ (3.9Kb positive), and have compared these with other R locus genes. From genetic and molecular analysis, it is suggested that R-mb complex consists two copy of R elements, and each element shows the paramutagenic and gene silencing effects by the fashion of cis-inactivation.

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Mapping of RFLP Markers Linked to Bacterial Blight Resistant Genes (Xa-1, Xa-3) in Rice (벼 흰잎마름병 저항성 유전자(Xa-1, Xa-3)연관 RFLP 마커 탐색)

  • 강현중;김현순;남정권;이영태;이승엽;김석동
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.6
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    • pp.419-423
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    • 2003
  • Bacterial blight caused by Xantomonas oryzae pv. oryzae is one of the most serious diseases of rice especially in southern area of Korea. Three races, $\textrm{K}_1$, $\textrm{K}_2$ and $\textrm{K}_3$, are the most dominant species. lo improve rice breeding efficiency using marker assisted selection, some RFLP markers were surveyed for polymorphism between resistant and susceptible to $\textrm{K}_1$ and $\textrm{K}_3$. And, 127 doubled-haploid (DH) lines derived from Milyang121/HRl1650-1-4-2 and 131 DH lines derived from Milyang123/HR10624-AC5 were evaluated to bacterial blight ($\textrm{K}_1$ and $\textrm{K}_3$). Milyang121 and HR10624-AC5 have Xa-1, resistant to $\textrm{K}_1$ race, and Milyang123 has Xa-3, resistant to $\textrm{K}_1$ and $\textrm{K}_3$ race. Three markers, RZ590, RZ536 and RG303, showing polymorphism between parents and resistance gene, Xa-1 and Xa-3, were analysed in the two combinations of DH lines. The segregation pattern of resistant DH population of Milyang123/HR10624-AC5 to susceptible showed 3:1 and 1:1 in $\textrm{K}_1$ and $\textrm{K}_3$ race. In three RFLP markers, RZ590 was linked to Xa-1 on chromosome 4, and RZ536 and RG303 were linked to Xa-3 on chromosome 11. The map distance between Xa-1 and RZ590 was 3.1cM on chromosome 4, and Xa-3 and RZ536/RG303 were 7.6/16.0cM on chromosome 11, respectively. The results of RFLP mapping will be useful for the selection and pyramiding of bacterial blight resistant genes.

Expression and Regulatory Analysis of Sporulation Gene (spo 5) in Schizosaccharomyces pombe (Schizosaccharomyces pombe 포자형성유전자 (spo 5)의 발현조절기구의 해석)

  • KIM Dong-Ju;SHIMODA Chikasi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.1
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    • pp.46-54
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    • 1997
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by malting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB (spo 5)1. We futher analyzed six recombinant plasmids, pDB (spo 5)2, pDB(spo 5)3, pDB(spo 5)4, pDB(spo 5)5, pDB(spo 5)6, pDB(spo 5)7, and found each plasmids is able to rescue the spo 5-2, spo 5-3, spo 5-4, spo 5-5, spo 5-6, spo 5-7, mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB (spo 5)1, and pDB (spo 5)R1 contained the spo 5 gene. Transcipts of spo 5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 25 kb were detected with 5 kb Hind III fragment containing a part of the spo 5 gene as a probe. The small mRNA (2.5 kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2 kb) was produced constitutively. Appearance of a 2.5 kb spo 5-mRNA depends upon the function of the mei1, mei2 and mei3 genes.

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Physical Environmental Factors Affecting Job Satisfaction - Focused on Office Environment of Oklahoma State University′s Faculty Members- (직업 만족도에 영향을 미치는 물리적 환경 요소들 - 오클라호마 주립대학 교수의 연구환경을 중심으로 -)

  • Hye-Sun, Han
    • Korean Institute of Interior Design Journal
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    • v.13 no.5
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    • pp.206-214
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    • 2004
  • 본 연구는 사무공간에 영향을 주는 물리적 환경 요소 (공간, 가구배치, 미, 주변요소)를 제시하고 각 환경 요소들과의 관계를 파악하며 각 물리적 환경요소들과 직업 만족도 (급여, 진급, 책무, 상사와의 관계, 물리적 공간과 배치, 수업방법의 향상, 현재 직업 만족도)와의 관계를 분석하는데 연구의 목적이 있다. 또한 물리적 환경 요소들, 직업 만족도와 개인 신상 (나이, 성별, 지위, 교육수준, 교육 경력) 의 특성들과의 상관관계를 알아보고자 한다. 본 연구는 오클라호마 주립대학교의 종신 재직권을 가진 교육대학 교수들을 대상으로 세 가지 가설을 바탕으로 작성된 설문지를 배포하여 조사되었다. 각 교수들의 사무공간은 1997년에 개보수를 마치고 그 해 여름부터 사용하고 있다. 설문은 현재 사용하고 있는 사무공간에 대한 평가, 직업 만족도에 대한 평가와 개인 신상의 특성에 대한 질문으로 조사되었다. 조사 결과 사무공간에 영향을 주는 물리적 환경 요소들 간에 밀접한 관계가 있다는 가설이 입증되었고, 또한 조사 결과 물리적 환경의 만족도가 증가할수록 직업만족도가 증가하는 것으로 물리적 사무환경에 대한 만족도와 직업에 대한 만족도가 직접적인 관계가 있다는 가설 또한 확증되었다. 물리적 환경요소, 직업 만족도와 개인 신상의 특성들과의 상관관계가 있다는 가설은 전체적으로는 입증되지는 않았지만 부분적으로 요소들 간에 관계를 보여주고 있다. 남성이 여성보다 가구배치에 대해 불만족스러웠고 교육경력이 적을수록 사무공간의 냉난방과 환기에 대한 불만족을 보였다. 본 연구의 결과는 교수들의 사무환경의 부정적, 긍정적 견해를 지각하여 사무 환경의 질을 높이는데 도움이 되는데 중요한 역할을 할 수 있겠다. 본 연구는 교육대학의 교수들에 국한되어 있었던 바 추후에는 전체 교육자들을 대상으로 장기간의 관찰을 통한 연구가 요구되어 진다.신만의 고유한 해결책으로 발전시키는지를 고찰하고자 한다. 본 논문의 목적은 서울을 사례분석 중 하나로 사용하여 인테리어 디자인 분야의 경계를 대도시로 확장하는 동시에 새로운 적용영역의 가능성을 탐구하는데 있다 하겠다.[C/N]의 값을 나타내었다.다.다.화 기술, 구동방법등에 대한 기술개요와 국내외 기술동향에 대하여 소개하고자 한다.었다.다._{2}$가 0.25[wt%] 첨가된 시편의 20[.deg.C]에서의 유전상수는 16,700으로 최대값을 유전손실을 1.28[%]로 최소값을 나타내었다. 또한 모든 시편은 온도 및 주파수에 따라 유전상수가 완만하게 변화하는 유전이완 특성을 나타내었다.다.수적인 물의 양에 따른 DIAION WA30의 라세미화 효율에 관하여 실험한 결과, 물의 양이 증가할수록 그 효율은 감소하였다. DIAION WA30을 라세미화 촉매로 사용하여 아이소옥탄 내에서 라세믹 나프록센 2,2,2-트리플로로에틸 씨오에스터의 효소적 DKR 반응을 수행해 보았다. 그 결과 DIAION WA30을 사용하지 않은 경우에 비해 반응 전환율과 생성물의 광학 순도는 급격히 향상되었다. 전통적 광학분할 반응의 최대 50%라는 전환율의 제한이 본 연구에서 찾은 DIAION WA30을 첨가함으로써 성공적으로 극복되었다. 또한 고체 염기촉매인 DIAION WA30의 사용은 라세미화 촉매의 회수 및 재사용이 가능하게 해준다.해준다.다. TN5 세포주를 0.2 L 규모 (1 L spinner flask)oJl에서 세포간의 응집현상 없이 부유배양에 적응,배양시킨 후 세포성장 시기에 따른 발현을 조사한 결과 1 MOI의 감염조건 하에서는 $0.6\times10^6$cell/mL의 early exponential시기의 세포밀도에서 72시간 배양하였을 대 최대 발현양을 나타내었다. 나타내었다.

Loss of Specific Sequences in a Natural Variant of Potato Proteinase Inhibitor II Gene Results in a Loss of Wound-Inducible Gene Expression (감자의 단백질 분해효소 억제제 II 유전자의 특별한 염기서열의 자연적 제거로 인한 상처 유발성 발현의 소실)

  • Thornburg, Robert W.;Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.104-111
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    • 1996
  • We have isolated several proteinase inhibitor II genes pin2 from a Russet Burbank potato DNA library. One of these, pin2T was subcloned and a 1.8 kb Xbal/Nsil insert was sequenced. This fragment contained the complete Inhibitor II gene including 965 Up of flanking DNA upstream from the gene and 200 bp of flanking DNA downstream from the gene. The open reading frame encodes a protein that is similar to other reported proteinase Inhibitor II proteins. The DNA sequence of the 5' flanking region of pin2T from -714 to +1 is highly homologous (91% identity) with that of the previously isolated wound-inducible pin2K. There are, however, four small deletions in the pin2T promoter which are located at -221 to -200, -263 to -254, -523 to -426 and -759 to -708 relative to the transcription start site of the wound-inducible pin2K. Three of these deletions map to a portion of the promoter that controls the wound-inducibility of the proteinase inhibitor genes. Chimeric genes containing the promoter of the pin2T gene linked with the both CAT and GUS were constructed and transfered into tobacco plants. Analysis of these plants indicated that pin2T is not a wound-inducible gene but is expressed at low levels. Thus, wound-inducibility is lost with the concomitant natural deletion of three small regions of the promoter. Comparision of the sequences deleted in pin2T relative to the pin2K with Genebank sequences indicates that the deleted sequences contain a motif (consensus 5'-AGTAAA-3') that is found in many other wound-inducible genes but not easily found in the published promoter sequences of other plant genes. Nuclear proteins from unwounded and wounded potato leaves were bound to the proximal promoter region, downstream of the 5'-AGTAAA-3', of pin2T. The comparison of the pin2T gone with the pin2K gene indicates that the natural internal promoter deletions are likely responsible for loss of the wound-inducible phenotype in the pin2T gene.

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Cloning and Transcription Analysis of Sporulation Gene (spo5) in Schizosaccharomyces pombe (Schizosaccharomyces bombe 포자형성 유전자(spo5)의 Cloning 및 전사조절)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.112-118
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    • 2002
  • Sporulation in the fission yeast Schizosaccharomyces pombe has been regarded as an important model of cellular development and differentiation. S. pombe cells proliferate by mitosis and binary fission on growth medium. Deprivation of nutrients especially nitrogen sources, causes the cessation of mitosis and initiates sexual reproduction by matting between two sexually compatible cell types. Meiosis is then followed in a diploid cell in the absence of nitrogen source. DNA fragment complemented with the mutations of sporulation gene was isolated from the S. pombe gene library constructed in the vector, pDB 248' and designated as pDB(spo5)1. We futher analyzed six recombinant plasmids, pDB(spo5)2, pDB(spo5)3, pDB(spo5)4, pDB(spo5)5, pDB (spo5)6, pDB(spo5)7 and found each of these plasmids is able to rescue the spo5-2, spo5-3, spo5-4, spo5-5, spo5-6, spo5-7 mutations, respectively. Mapping of the integrated plasmid into the homologous site of the S. pombe chromosomes demonstrated that pDB(spo5)1, and pDB(spu5)Rl contained the spo5 gene. Transcripts of spo5 gene were analyzed by Northern hybridization. Two transcripts of 3.2 kb and 2.5kb were detected with 5kb Hind Ⅲ fragment containing a part of the spo5 gene as a probe. The small mRNA(2.5kb) appeared only when a wild-type strain was cultured in the absence of nitrogen source in which condition the large mRNA (3.2kb) was produced constitutively. Appearance of a 2.5kb spo5-mRNA depends upon the function of the meil, mei2 and mei3 genes.