• Title/Summary/Keyword: 제한 효소 지도 작성

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Development of Restriction Fragment Length Polymorphism(RELP) Markers in Silkworm, Bombyx mori (누에 RFLP(제한단편 다형현상)마커 개발)

  • 고승주;김태산;이영승;황재삼;이상몽
    • Korean journal of applied entomology
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    • v.36 no.1
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    • pp.96-104
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    • 1997
  • A silkworm Bombyx mori genomic DNA library was constructed from polyphagous J111 strain and unpolyphagous $C_3$ strain to develop the genomic study by DNA makers. Genomic DNAs of two strains were digested with restriction enzyme EcoRI and ligated into pUC18. The ligated plasmids were transferred into E. coli host strain DH5$\alpha$. When the genomic DNAs were hybridized with insert DNAs from transformant, could be categorized from hybridization patterns to three groups as high repetitive sequence, moderately repetitive sequence, and low-copy number sequences. A total of 219 clones containing single or low-copy number sequence inserts were examined for any polymorphisms between two strains of J111 and $C_3$. Forty six clones showed RFLPs and 10 of these clones were used as a probe of analysis of $F_2$ population derived from crossing between J111 and $C_3$ strain. The genetic inheritance tested with each clones will be important tools to construct the genetic map of the silkworm, Bombyx mori.

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Mapping of UV-B sensitive gene in Arabidopsis by CAPS markers (CAPS marker에 의한 Arabidopsis의 자외선 B 감수성 유전자 지도작성)

  • 박홍덕;김종봉
    • Journal of Life Science
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    • v.12 no.6
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    • pp.715-720
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    • 2002
  • A mutant Arabidopsis thaliana which is very sensitive to Ultraviolet-B(UV-B) radiation has been isolated by ethylmethane sulfonate(EMS) mutagenesis. Genetic cross proved the UV-sensitive gene(uvs) to segregate as a single Mendelian locus. For mapping of uvs, we crossed Arabidopsis thaliana Lansberg with uvs plant(Columbia), and made F2 plants by F1 selfcross. We designed 10 kinds of CAPS marker primers. Each primers amplifies a single mapped DNA sequence from uvs and Lansberg erecta ecotyres. Also identified was at least one restriction endonuclase for each of these PCR product that generates ecotype-specific digestion pattern. We got crossing over value of UB-sensitivity and each CAPS marker which located on different chromosome arm. The value of crossing over showed that uvs was linked to LFY3 which was on chromosome 5.

Genetic Structure of the phnM Gene Encoding Plant-Type Ferredoxin from Pseudomonas sp. strain DJ77 (Pseudomonas sp. strain DJ77에서 Plant-Type의 Ferredoxin을 암호화하는 phnM 유전자의 구조)

  • Kim, Sungje;Kim, Young-Chang
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.115-119
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    • 1998
  • We cloned the 4.8 kb BglII fragment containing genes downstream pHENX7 from Pseudomonas sp. strain DJ77. The restriction map of the resultant clone, recombinant plasmid pYCS500, was determined. Sequencing analysis of the 465 bp HindIII-ClaI fragment revealed an open reading frame of 282 bp that was then designated phnM. The deduced polypeptide is 93 amino acid residues long with a $M_r$ of 10,008. The PhnM has 37.3-53.9% identity with plant-type ferredoxin proteins such as NahT, XylT, DmpQ, AtdS, PhlG, PhhQ and TbuW and contains the motif similar to well-conserved functional domains of those proteins.

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Complementary DNA Cloning and Restriction Mapping of Nuclear Inclusion Body and Coat Protein Genes of Turnip Mosaic Virus-Ca Strain Genomic RNA (순무모자이크 바이러스 Ca계통 핵봉입체와 외피단백질 유전자의 cDNA 클로닝 및 제한효소 지도작성)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.235-239
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    • 1994
  • Viral RNA was extracted from purified Chinese cabbage strain of turnip mosaic virus (TuMV-Ca) from infected leaves of turnip. Polyadenylated genomic viral RNA was recovered by oligo (dT) cellulose column chromatography and used as a template for the synthesis of complementary DNA (cDNA). Recombinant plasmids contained cDNA ranged from about 900 bp to 2, 450 bp were synthesized. Among the selected 41 transformants, pTUCA31 and pTUCA35 had over 2 Kbp cDNA insert. Restriction endonuclease patterns of the clones examined were very similar among them. Clones pTUCA23 and pTUCA31 were overlapped with pTUA35. The longest clone pTUCA35, encoding 3'-end, showed that it contained two sites for EcoRI, and one site for BamHI, ClaI, HincII, SacI and XbaI, respectively. The restriction mapping indicated that the clone pTUCA35 contained partial nuclear inclusion body gene, complete coding region of the coat protein and 3' untranslated region of TuMV-Ca genomic RNA.

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Characterization of Spodoptera exigua Nuclear Polyhedrosis Virus Polyhedrin Gene Structure (파밤나방 핵다각체병 바이러스의 다각체 단백질 유전자 구조)

  • 최재영;김우진
    • Journal of Sericultural and Entomological Science
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    • v.38 no.2
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    • pp.144-149
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    • 1996
  • To develope the baculovirus expression vector system (BEVS) using Spodoptera exigua nuclear polyhedrosis virus (SeNPV), we characterized the polyhedrin of SeNPV. The SeNPV polyhedra was irregular and composed of the major protein molecular weight of 30 kDa determined by electronmicroscopy and SDS-AGE analysis, respectively. The nucleotid suquences of 876 bases including the coding region of polyhedrin gene was determined and it was revealed that the polyhedrin gene is located within Xho I 3.0Kb and Nco I 6.0 Kb by Southern blot analysis, respectively. Also, the Xho I 3.0 Kb and the Nco I 6.0 Kb fragments were cloned and restriction enzyme map of these clones were determined.

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Restriction endonuclease maps of three plasmids from bacillus thuringiensis serovar israelensis 4Q1 (Bacillus thuringiensis serovar israelensis 4Q1로부터 분리된 plasmid 제한효소지도 작성)

  • Faust, R.M.;;;C.L.Meyers-Dowling;P.E.McCawley
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.122-128
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    • 1985
  • Bacillus thuringiensis serovar israelensis 4Q1 contains 8 different covalently closed-circular (CCC) plasmids of molecular weight 204, 267, 109, 103, 16, 7.6, 6.4, and 5.0kb. The three smallest plasmids, designated pBti6, and pBti8 may prove to be useful as cloning vectors because of thier size and ease of isolation. The three plasmids were incubated separately with 9 different restriction enzymes and 7 of the enzymes tested cleaved one or more of the plasmids. Plasmid pBti6 has a single site for Bg1 II, Pst I and Pvu II, two sites for Bc1 I and Eco RI, and five sites for Hind III. Plasmid pBti7 has a single site for Bam HI and Pst I, two sites for Hind III, and three sites for PvuII. Plasmic pBti8 has a single site for Bam HI, BelI and Hind III, two sites for Eco RI, and three sites for Bgl II and Pvu II. Composite restriction enzyme maps for pBti6, pBti7 and pBti8 were constructed. The sites of restriction enzyme cleavage were determined by single, double and partial digests of the plasmid DNA. All the restriction sites were aligned relative to the single Bgl II(pBti6), Pst I(pBti7) or Hind III(pBti8) site, respectively.

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Molecular Cloning of Bacteriocin Gene and Biological Control of Plant Pathogen (Bacteriocin 생산 유전자의 Cloning 및 식물병원균에 대한 생물학적 억제)

  • 김교창;육창수;도대홍
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.98-102
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    • 1990
  • A strain of Erwinia spp. was selected from the soil for the production of bacteriocin to the root rot plant pathogen. Bacteriocin producing gene was not located on plasmid but on chromosome. Genomic library of Erwinia spp. were made by using pLAFR 3 as a vector system for cloning of the gene. It was been cloned and expressed in E. coli DH 5 . Bacteriocin producing colony was composed of pLAFR 3 vector and 3.0 kb EcoRI fragment of Erwinia spp. ehromosomal DNA. The inserted fragment (3.0 kb) was possessed a EcoRI and BarnHI restriction sites.

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Incorporation of RAPD linkage Map Into RFLP Map in Glycine max (L, ) Merr (콩의 RAPD 연관지도를 RFLP 연관지도와 합병)

  • Choi, In-Soo;Kim, Yong-Chul
    • Journal of Life Science
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    • v.13 no.3
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    • pp.280-290
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    • 2003
  • The incorporation of RAPD markers into the previous classical and RFLP genetic linkage maps will facilitate the generation of a detailed genetic map by compensating for the lack of one type of marker in the region of interest. The objective of this paper was to present features we observed when we associated RAPD map from an intraspecific cross of a Glycine max$\times$G. max, 'Essex'$\times$PI 437654 with the public RFLP map developed from an interspecific cross of G. max$\times$G. soja. Among 27 linkage groups of RAPD map, eight linkage groups contained probe/enzyme combination RFLP markers, which allowed us the incorporation of RAPD markers into the public RFLP map. Map position rearrangement was observed. In incorporating L.G.C-3 into the public RFLP linkage group a1 and a2, both pSAC3 and pA136 region, and pA170/EcoRV and pB170/HindIII region were in opposite order, respectively. And, pk400 was localized 1.8 cM from pA96-1 and 8.4 cM from pB172 in the public RFLP map, but was localized 9.9 cM from i locus and 18.9 cM from pA85 in our study. A noticeable expansion of the map distances in the intraspecific cross of Essex and PI 437654 was also observed. Map distance between probes pA890 and pK493 in L.G.C-1 was 48.6 cM, but it was only 13.3 cM in the public RFLP map. The distances from the probe pB32-2 to pA670 and from pA670 to pA668 in L.G. C-2 were 50.9 cM and 31.7 cM, but they were 35.9 cM and 13.5 cM in the public RFLP map. The detection of duplicate loci from the same probe that were mapped on the same or/and different linkage group was another feature we observed.

Studied on Amplificative Efficiency of PCR of Predigested template DNA and GC Contents for RAPD Analysis in the Silkworm, Bombyx mori (누에의 RAPD 분석을 위한 primer의 GC 함량과 사전 제한효소 처리한 주형 DNA의 PCR 증폭효율에 관한 연구)

  • 이진성;황재삼;이상몽;황석조;강현아;성승현;서동상
    • Korean journal of applied entomology
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    • v.35 no.1
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    • pp.58-65
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    • 1996
  • This experiment was carried out to evaluate the effect of sublethal doses of BPMC, etofenprox, and buprofezin on N. lugens. and its predator C. lividipennis. Buprofezin was found to be the most toxic to N. lugens and the most safe to C. lividipennis among the three insecticides, based on LD50 values. Selective toxicity index calculated by dividing LDSo value of C. lividipennis by that of N. lugens indicated that buprofezin was very safe to C. lividipennis, showing selective toxicity of 2703.3. Longevity and fecundity of N. lugens treated with LDIU and LDm of buprofezin and BPMC were not significantly different with those of untreated brown planthoppers. However, egg hatchability' of N. lugens was greatly reduced when treated with LDm of buprofezin, having the highest inhibition rate of 17.7%. Hatchability of eggs from insects treated with BPMC was similar to that of control. The oviposited peak of treated hoppers appeared late as compared to the untreated which showed the peak at early part of the ovipositional period. The longevity and fecundity of C. lividipennis treated with BPMC were significantly reduced as compared with the untreated. Etofenprox also induced fecundity reduction when treated with LDlo, and LDm. However, C. lividipennis treated with sublethal doses of buprofezin showed no redution in logevity and fecundity. From these results, it may be said that buprofezin can be used to control brown planthopper without disrupting of C. lividipennis population in the rice field.

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Characterization of a Restriction Endonuclease, SdiI from Streptomyces diastatochromogenes (Streptomyces diastatochromogenes로부터 분리된 SdiI의 특성에 관한 연구)

  • Bae, Moo;Song, Eun-Sook;Hwang, Hye-Yeon;Yim, Jeong-Bin
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.301-305
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    • 1994
  • In catalytic properties of the restriction enonuclease, SdiI, which was purified from Streptomyces diastatochromogenes, this enzyme was active at wide range between pH 7.0 and 12.5, and up to $60^{\circ}C$ and 500 mM of NaCl concentration. It was stable between 20^{\circ}C$ and $60^{\circ}C$, and essentially requires $MgCl_2$ for endonuclease activity. The restriction map of lambda DNA which was obtained by double digestion with various enzymes suggested SdiI to be an isoschizomer of XhoI. From the determination of restriction site based on DNA sequencing method, recognition and cleavage specificity of SdiI was concluded as: 5‘-C${\downarrow}$TCGA G-3' 3'-G AGCT${\uparrow}$C-5'

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