• Title/Summary/Keyword: 제한 효소 지도 작성

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Construction of Genetic Linkage Map and Identification of Quantitative Trait Loci in Populus davidiana using Genotyping-by-sequencing (Genotyping-by-sequencing 기법을 이용한 사시나무(Populus davidiana) 유전연관지도 작성 및 양적형질 유전자좌 탐색)

  • Suvi Kim;Yang-gil Kim;Dayoung Lee;Hye-jin Lee;Kyu-Suk Kang
    • Journal of Korean Society of Forest Science
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    • v.112 no.1
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    • pp.40-56
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    • 2023
  • Tree species within the Populus genus grow rapidly and have an excellent capacity to absorb carbon, conferring substantial ability to effective purify the environment. Poplar breeding can be achieved rapidly and efficiently if a genetic linkage map is constructed and quantitative trait loci (QTLs) are identified. Here, a high-density genetic linkage map was constructed for the control pollinated progeny using the genotyping-by-sequencing (GBS) technique, which is a next-generation sequencing method. A search was also performed for the genes associated with quantitative traits located in the genetic linkage map by examining the variables of height and diameter at root collar, and resilience to insect damage. The height and diameter at root collar were measured directly, while the ability to recover from insect damage was scored in a 4-year-old breeding population of aspen hybrids (Odae19 × Bonghyeon4 F1) established in the research forest of Seoul National University. After DNA extraction, paternity was confirmed using five microsatellite markers, and only the individuals for which paternity was confirmed were used for the analysis. The DNA was cut using restriction enzymes and the obtained DNA fragments were prepared using a GBS library and sequenced. The analyzed results were sorted using Populus trichocarpa as a reference genome. Overall, 58,040 aligned single-nucleotide polymorphism (SNP) markers were identified, 17,755 of which were used for mapping genetic linkages. The genetic linkage map was divided into 19 linkage groups, with a total length of 2,129.54 cM. The analysis failed to identify any growth-related QTLs, but a gene assumed to be related to recovery from insect damage was identified on linkage group (chromosome) 4 through genome-wide association study.

Cloning of the Polyhedrin Gene-Containing DNA Fragment of Hyphantria cunea Nuclear Polyhedrosis Virus (흰불나방 핵다각체바이러스 다각체단백질 유전자포함 절편의 클로닝)

  • 박호영;진병래;박순식;김정일;깅석권
    • Korean journal of applied entomology
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    • v.32 no.1
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    • pp.51-60
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    • 1993
  • The polyhedrin gene-containing DNA fragment of Hyphantria cunea nuclear polyhedrosis virus (HcNPV) was localized by southern hybridization with Autographa california CPA EcoRI-I fragment (7.3 kb), Bombyx mori NPV PatI-F fragment (7 kb) and synthetic oligonucleotide(30-mer) as probes. the PstI-L(5.3 kb) fragment of HcNPV was cloned to E. coli and the plasmid of the fragment was named as pHcP-L(8.0 kb). The pHcP-L was physically mapped and subcloned to E. coli as pHcP-L1(4.7 kb), pHcP-L2(7.1 kb), pHcP-L3(5.3 kb), pHcP-L4(4.2 kb) and pHcP-L5(4.5 kb).

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Molecular Cloning and Expression of Human Poly (ADP-ribose) Synthetase cDNA in E. Coli (인간 Poly(ADP-ribose) Synthetase cDNA의 클로닝 및 대장균에서의 발현)

  • 이성용;김완주;이태성;박상대;이정섭;박종군
    • The Korean Journal of Zoology
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    • v.39 no.3
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    • pp.248-256
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    • 1996
  • The present study was performed to clone and express human poly (ADP-ribose) synthetase (PARS) cDNA in E coli. For these purposes, the CDNA for human poly (ADP-ribose) synthetase, encoding the entire protein, was cloned into pGEM-7Zf(+). The resulting recombinant plasmid pPARS6.1 was restriction enzyme mapped and its identity was confirmed by Southern blot analysis. The pPARS6. 1 contained full-length CDNA of human PARS and the nudeotide sequences were identical with those reported previously. The recombinant protein which migrated as a unique 120 kDa band on 10% SDS-polyacrylamide gels, was identified as PARS by Southwestern blots using nick-translated DNA probes and by activity gels and activity blots using 32 P-NAD as a substrate for poly (ADP-ribose) synthetase (PARS). The signals corresponding to 120 and 98 kDa proteins were obtained following IPTG (0.4 mM) induction of the PARS cDNA cloned into Xba I-digested pGEM-7Zf(+) vector. Nonspecific signals corresponding to 45 and 38 kDa proteins were also shown in both IPTG-induced and noninduced cells. The nonspecific proteins may be products of incomplete translation or proteolytic products of intact PARS.

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Location and Nucleotide Sequence of the Bombyx mori Nuclear Polyhedrosis Virus Polyhedrin Gene (누에 핵다각체병 바이러스의 다각체 단백질 유전자의 위치 탐색 및 염기서열)

  • 우수동;김현욱;박범석;강석권;양재명;정인식
    • Journal of Sericultural and Entomological Science
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    • v.34 no.2
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    • pp.20-25
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    • 1992
  • The location of the polyhedrin gene of Bmbyx mori nuclear polyhedrosis virus(BmNPV) was determined by using a cloned polyhedrin gene from the Autographa californica nuclear polyhedrosis virus(AcNPV) as a hybridization probe. The 7.4 Kb PstⅠ fragment DNA of Bm-NPV was cloned to plasmid pUC19 vector. A fragment containing this gene was mapped and sequenced in its entire polyhedrin reading frame. Nucleotide sequences comparison of the polyhedrin of the BmNPV to that of previously reported by Ⅰatrou(1985) revealed that the sequence varied in 10 base, Comparison of the amino acid sequence of the two structured gene revealed that coding sequence varied 74 valine to isoleucine, 76 aspargine to serine and 155 methionine to valine.

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Molecular Cloning of nifHD from Rhizobium sp. SNU003 (Rhizobium sp. SNU003의 nifHD 클로닝)

  • 강명수;안정선
    • Korean Journal of Microbiology
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    • v.31 no.2
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    • pp.123-128
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    • 1993
  • Genes for dinitrogenase reductase (nifH) and dinitogenase a subunit (nifD) were found to be located on 7.9 kb of EcoRI, 6.5 kb of Sail, 7.3 kb of HindlII and 4.4 kb of Pstl fragments of the genomic blot of Rhizobium sp. SNU003. a symbiotic strain from root nodule of Canavalia lineata. Nine recombinant phage nif-clones were selected from the genomic library constructed by using EMBL-3 BamHI arms of bacteriophage lambda. Among them. Rnif-6 had insert DNA of 15.3 kb. in which 7.6 kb of BamHI!SacI fragment contained nifHD region. Therefore, the 7.6 kb fragment was subcloned into pUC19 and partial restriction map was constructed. As the results, nifH and nifD were found to be located continuously on 4.5 kb of BamHI/BglIl in the genome of Rhizobium sp. SNU003 strain.

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Sequence Analysis, Molecular Cloning and Restriction Mapping of Mitochondreal Genome of Domesticated Silkworm, Bombyx mori (누에 미토콘드리아 유전체의 제한효소 지도작성, 클로닝 및 염기서열 분석)

  • 이진성;성승현;김용성;서동상
    • Journal of Sericultural and Entomological Science
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    • v.42 no.1
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    • pp.14-23
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    • 2000
  • The mitochondrial genome of domesticated silkworm (Bombyx mori) was mapped with five restriction endonucleases (BamHI, EcoRI, HindIII, PstI and XbaI), the entire genome was cloned with HindIII and EcoRI. From the end sequencing results of 5$^1$and 3$^1$region for full genome set of eleven mitochondrial clones, the seven mitochondrial genes (NADH dehydrogenase 6, ATPase 6, ATPase 8, tRN $A^{Lys}$, tRN $A^{Asp}$, tRN $A^{Thr}$ and tRN $A^{Phe}$ of mori were identified on the basis of their nucleotide sequence homology. The nucleotide composition of NADH dehydrogenase 6 was heavily biased towards adenine and thymine, which accounted for 87.76%. On basis of the sequence similarity with published tRNA genes from six insect species, the tRN $A^{Lys}$, tRN $A^{Asp}$ and tRN $A^{Thr}$ were showed stable canonical clover-leaf tRNA structures with acceptible anticodons. However, both the DHU and T$\psi$C arms of tRN $A^{Phe}$ could not form any stable stem-loop structure. The two overlapping gene pairs (tRN $A^{Lys}$ -tRN $A^{ASP}$ and ATPase8-ATPase6) were found from our sequencing results. The genes are encoded on the same strad. ATPase8 and ATPase6 overlaps (ATGATAA) which are a single example of overlapping events between abutted protein-coding genes are common, and there is evidence that the two proteins are transcribed from a single bicistronic message by initiation at 5$^1$terminal start site for ATPase8 and at an internal start site for ATPase6. Ultimately, this result will provide assistance in designing oligo-nucleotides for PCR amplification, and sequencing the specific mitochondrial genes for phylogenetics of geographic races, genetically improved silkworm strains and wild silkworm (mandarina) which is estimated as ancestal of domesticated silkworm.sticated silkworm.

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Construction of Plasmid Vectors for Zymomonas mobilis (Zymomonas mobilis의 Plasmid Vector 제조에 관한 연구)

  • Hwang, Duk-Ju;Rhee, Sang-Ki;Pack, Moo-Young
    • Microbiology and Biotechnology Letters
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    • v.15 no.5
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    • pp.319-327
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    • 1987
  • In order to develop useful plasmid vectors for Zymomonas cells, attempts were made to isolate natural plasmids from Z. mobilis ATCC10988. Among a few plasmids isolated, a small plasmid of 3.9 Kb size was chosen and designated as pZM3. By introducing the replication origin of pZM3 into pBR325, a hybrid plasmid vector of 8.4 Kb size, pHZ22, was constructed. This vector contained chloramphenicol resistant gene as a selectable marker and proved to be conjugally transmissible and stably maintained in Z. mobilis. Tetracycline resistant gene was isolated from RP4 and introduced into pHZ22 to make a new vector called pHZT224 of 10.7 Kb size. Through n series of experiments, it was evident that these plasmid vectors containing selectable markers of chloramphenicol and tetracycline resistance were shuttle vectors functional in Z. mobilis as well as E. coli.

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Characterization of plasmids of Zymomonas mobils and Construction of E. coli-Zymomonas shuttle Vector (Zymomonas mobilis플라스미드의 특성연구 및 E.coli-Zymomonas셔틀 벡터 제조)

  • 이용억;이병재;강현삼
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.56-63
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    • 1985
  • We have characterized the plasmids of zymomonas, and constructed E. coli-Zymomonas shuttle vector. Plasmids have been detected in four strains of Zymomonas mobilis. All strains tested had at least one plasmid ranging in size from about 1.7 to 46kb. Antibiotics resistances of Z. mobilis were tested to select the host strain. All strains were very sensitive to tetracycline and chloramphenicol. Homology tests between the plasmids in four strains showed that the plasmids of ATCC10988 is highly homologous to those of ZM1, and that there is no homology between plasmids of ZM4 and Agll. The 1.7kb plasmid of ATCC10988, named as pZM886, also has no homology with plasmids of ZM4. A hybrid plasmid, designated to pBZ41, was constructed from pZM886 and pBR322. A restriction map of pBZ41 was established. Replicon of pZM886 didn't operate in E.coli and pBR322 seemed not to replicate in Zymomonas. pBZ41 was transfered from E. coli to Zymomonas by conjugal mobilization. The transconjugants were resistant to tetracycline and maintained pBZ41 stably.

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Frankia sp. strain SNU 014201의 nif-H, D, K, 유전자 클로닝

  • 권석윤;강명수;안정선
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.30-36
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    • 1992
  • nif (nitrogen fixation)-H.D, K genes of Frankia sp. SNU 014201. a symbiotic strain isolated from root nodule of Alnus hirsura, were found to be located in the genome on 13.5 kb of EcoRI, 18.0 kb of BamHI, 10.5 kb of BglII and 4.5 kb of KpnI fragments. Using EMBL-3 BamHI arms of bacteriophage lambda. the genomic library was constructed. from which fourteen recombinant phage nif-clones were selected. Among them, Ahnif-I2 had insert DNA of 18 kb, in which 7.9 kb of BamHl fragment contained nif-H, D, K and 3.6 kb of HindlIl/KpnI had nif-H and partial -D. Therefore, the 7.9 kb and 3.6 kb fragments were subcloned and partial restriction maps were constructed. As the results, nif-F1, D.K genes were found to be located continuously on the 6.5 kb of HindII/BamHI and 5.2 kb of SalIIBamHI fragment in the genome of Frankia sp. SNU 014201.

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Structure of Spodoptera exigua Nucleopolyhedrovirus p10 Gene (파밤나방 핵다각체병 바이러스의 p10 유전자 구조)

  • 최재영;우수동;홍혜경;이해광;제연호;강석권
    • Korean journal of applied entomology
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    • v.38 no.2
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    • pp.145-149
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    • 1999
  • To develop the baculovirus expression vector system (BEVS) adopting p10 gene promoter of Spodoptera exigua nucleopolyhedrovirus (SeNPV), we characterized the p10 gene of SeNPV. The nucleotide sequence of 545 bases including the coding region of p10 gene was determined. Compared with the previously reported SeNPV p10 gene (Zuidema et al., 1993), 4 bases were different in the 5' and 3' flanking region but no difference was found in the coding region. The p10 gene was located within Xho I 1.5 Kb, Sph 1 2.4 Kb and Cla I 4.0 Kb fragments by Southern hybridization analysis. Also, the Sph I 2.4 Kb and the Cla I 4.0 Kb fragments were cloned and their restriction enzyme maps were determined.

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