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Genetic variance of Tuchomonns uaginclis isolates by Southern hybridization (Southern hybridization에 의한 질편모충의 유전학적 다양성)

  • 류재숙;민득영
    • Parasites, Hosts and Diseases
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    • v.36 no.3
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    • pp.207-212
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    • 1998
  • In the present study, genomic DNAs were purified from Korean isolates (KT8, KT6, KT-Kim and KT-Lee) and foreign strains (CDC85, IR78 and NYH 286) of 1Trichomonas voslnalis, and hybridized with a probe based on the repetitive sequence cloned from T. uqfinolis to observe the genetic differences. By Southern hybridization, all isolates of T. uoSinoLis except the NYH386 strain had 11 bands. Therefore all isolates examined were distinguishable into 3 groups according to their banding patterns; i) KT8, KT6 and KT-Kim isolates had 11 identical bands such as 1 kb, 1.2 kb, 1.6 kb, 1.9 kb, 2.3 kb. 27 kb, 3.2 kb, 2.4 kb, 3.8 kb, 4.9 kb and 6.0 kb, ii) The metronidazole-resistant IR78 strain had the some bands as KT-Lee isolate at bands of 1 kb, 1.2 kb, 1.6 kb. 1.8 kb, 2.1 kb, 2.5 kb, 2.7 kb, 2.9 kb, 3.4 kb, 5.0 kb and 6.0 kb, Bands of CDC85, metronidazole-resistant strain, were similar to those of IR78 and KT-Lee, except that 3.2 kb replaced 2.9 kb. iii) NYH286 particularly had 12 bands and bun patterns were similar to IR78 with a few exceptions as follows; i) 6.2 kb in place of 6.0 kb, ii) 2.0 kb and 2.2 kb instead of 2.1 kb. Through the results obtained, genetic variance of T. uoginnlis isolates was demonstrated by Southern hybridization.

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Polymorphism of Carp $\beta$-globin Gene on Chromosome (잉어 $\beta$-globin 유전자의 염색체상에서의 다형해석)

  • 진덕희;청목주
    • Journal of Life Science
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    • v.8 no.3
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    • pp.348-351
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    • 1998
  • Common DNA fragments of the ${\beta}$-globin gene were observed from six races of the adult common carp: Hybrid-Yamato, Japanese wild type, Mirror, Suwa-Yamato, Scale German, and Saku-Yamato. Chromosomal DNAs isolated from the above six races were digested with restriction endonucleased EcoRI and PstI. The digested fragments were transferred onto nitrocellulose filter and hybridized with a probe of carp ${\beta}$-globin cDNA. Molecular sizes of the hybridized DNA fragments digested with EcoRI were 3.6Kb(Kilo base), 4.3Kb and 15Kb in Hybrid-Yamato, Japanese wild type, Mirror, Scale German and Saku-Yamato carp DNAs. In Scale German and Saku-Yamato carp DNAs, two and one more hybridized DNA fragments were observed, respectively. Molecular sizes of the hybridized DNA fragments digested with PstI were 2.2Kb, 6.5Kb, 7.8Kb and 9.2Kb in Hybrid-Yamato, 2.2Kb, 6.5Kb and 9.2Kb in Japanese wild type, 2.2Kb, 6.5Kb, 7.8Kb, and 13Kb in Mirror, 2,2Kb, 5,5Kb, 6.5Kb, 7.8Kb, 9.2Kb and13Kb in Scale German, and 2.2Kb, 5.5Kb, 6.5Kb, 9.2Kb and Saku-Yamato carp DNA. Therefore, depending on carps, three to six DNA fragments were hybridized with ${\beta}$-globin gene probe. Thus it indicated polymorphysm in the globin gene family of carp.

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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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Nucleotide Sequence of 7.2 kb Mitochondrial Linear Plasmid DNA in Pleurotus ostreatus (Pleurotus ostreatus 미토콘드리아의 7.2 kb 선상 플라스미드 염기서열 분석)

  • 윤혜숙;구용범;노정혜
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.37-41
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    • 2001
  • Two linear plasmid-like DNAs, 10.2 kb and 7.2 kb were found in the mitochondria of P. ostreatus. They have covalently linked 5'-terminal proteins in both ends. Two continuous fragments of 4.7 kb and 2.3 kb from 7.2 kb DNA were cloned and sequenced. Two long open reading frames (ORF1; 2982 bp, 993 a.a and ORF2; 2703 bp, 900 a.a) and one short open reading frame(ORF3; 771 bp, 256 a.a) were found in the 7.2 kb plasmid. The putative ORF1 and ORF2 have conserved motifs of DNA polymerases and RNA polymerases, respectively, while the ORF3 has homologous regions with phosphatase from Plasmodium, and also with adhesine from Mycoplasma.

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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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Development of Black Shank Resistant Burley Tobacco Germplasm, KB 104 and KB 106, and Their Characteristics (버어리종 역병저항성 계통 KB 104와 KB 106의 육성경과 및 특성)

  • 조천준;김대송;정석훈;최상주;조명조
    • Journal of the Korean Society of Tobacco Science
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    • v.17 no.1
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    • pp.57-61
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    • 1995
  • Black shank(Phytophthora parasitira roar. nicotianae) resistant burley tobacco (Nicotiana tabacum L.) germplasms, KB 104 and KB 106, were developed by Korea Ginseng & Tobacco Research Institute. KB 104 was developed from the single cross of Burley 21$\times$Newton 77, using a modified pedigree method. KB 104 was highly resistant to black shank, and its agronomic characteristics and chemical contents were comparable to those of Burley 21, and value per 10a was slightly higher than Burley 21, KB 106 is a maternally derived doubled haploid made by N. africana method from the single cross of Burley 21$\times$ Va 509. KB 106 was also highly resistant to black shank, had two more harvestable leaves per plant and flowered three days later than Burley 21 did. Total alkaloid and nicotine contents of KB 106 were significantly lower than those of Burley 21. But its nornicotine content was higher than Burley 21 5. Key wads : Burley tobacco germplasm, Black shank resistance.

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Effects of Sodding and Seeding Time and Rate of Seed Mixture on the Establishment of Cool-Season Turfgrasses (한지형 잔디의 조성시기, 초종 혼합 비율이 잔디 피복에 미치는 영향)

  • Shim Gyu Yul;Kim Chang Soo;Lee Seong Ho;Joo Young Kyoo
    • Asian Journal of Turfgrass Science
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    • v.18 no.4
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    • pp.179-191
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    • 2004
  • This study was conducted to find out the effect of sodding and seeding time and rate of seed mixtures on the establishment of cool-season turfgrasses by evaluating the turf coverage rates for two years. In fall planting, the required establishment period of full coverage($100\%$) was 1.5 months with a rolled turf sodding(Kentucky bluegrass $100\%$, Kentucky bluegrass $80\%$+perennial ryegrass $20\%$). The $100\%$ turf establishment was achieved in 7 months with Perennial ryegrass $100\%$, and 7.5 months by seeding with Kentucky bluegrass $100\%$(KB 100), Kentucky bluegrass $80\%$+perennial ryegrass $20\%$(KB80+PR20), Kentucky bluegrass $70\%$+perennial ryegrass $30\%$(KB70+PR30). In spring planting, the establishment periods far sod with KB 100 or KB80+PR20 were taken one month. However, in the case of seeding, the establishment periods were 3 months, 3.5 months, 3.5 months and 4 months with PR100, KB80+PR20, KB70+PR30, and KB 100, respectively Comparing the turf establishment vigor between fall and spring planting, the vigor was higher In spring planting than in fall planting in both sodding and . seeding. In the case of spring planting, the most proper time for turf establishment was tested on April, May, and June trials. The effect was significant in establishment vigor. The result showed highest on April planting. On May and June trials, establishment vigors were decreased gradually As the mixture rate of PR increased, ryegrass, establishment vigor was decreased with the rates. These results indicated that perennial ryegrass has relatively less tolerant to summer heat than Kentucky bluegrass. Number of shoots in 95 days after seeding was higher in KB100 by 16,600 per $m^2$ than in PR100 by 12,400 per $m^2$, while the lowest number showed in KB50+PR50 by 3,300 per $m^2$. Those in KB80:PR20, KB70:PR30 were 6,700 and 4,900 per $m^2$, respectively. The ratios of tillers according to mixture rates between Kentucky bluegrass and perennial ryegrass were KB80:PR20=87:13, KB70:PR30=78:22, and KB50:PR50=48:52. According to results in this study, Ideal seeding time might be spring (April) than in fall (September), and proper mixture rate was $80\%$ of Kentucky bluegrass with $20\%$ of perennial ryegrass.

형질전환생쥐에서 1.7 kb 및 3.1 kb bovine $\beta$-casein promoter가 human type II collagen 유전자의 발현조절에 관한 분석

  • 나루세겐지;양정희;권혁빈;유승권;최윤재;박창식;진동일
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.89-89
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    • 2003
  • 본 연구에서는 1.7kb 및 3.1kb bovine $\beta$-casein promoter의 유전자 발현 조절능력을 알아보기 위해 1 7kb 및 3.1kb bovine $\beta$-casein promoter에 human Type II Collagen 유전자를 연결해서 DNA microinjection으로 형질전환생쥐를 생산하였다. 총 8마리의 founder생쥐(1.7kb collagen : 5마리, 3.1kb collagen 3마리)를 생산하였고 이 founder생쥐와 wild type 생쥐를 mating시켜서 $F_1 및 F_2$ 새끼를 얻었다. $F_1 및 F_2$새끼들에서 human Type II collagen 유전자의 transmission rate는 약 50%로 Mendel의 법칙에 따라 분리되어 안정적으로 유전자가 염색체에 정착되어 있음을 확인하였다. 이들 $F_1 및 F_2$새끼 중 암컷들을 임신시켜 분만 후 5-10 일경에 유선조직을 포함하여 여러 조직으로부터 RNA를 추출하여 Northern blotting 및 RT-PCR 방법을 이용하여 Type II collagen mRNA의 발현을 분석하였다. 유선에서의 발현은 1 7 kb 및 3.1 kb line별로 각각 1 line씩 발현되지 않았고, 그 외 line에서는 모두 발현되는 것으로 확인되었다. 유선에서의 Type II collagen mRNA 발형양은 1.7 kb 및 3.1 kb bovine $\beta$-casein promoter사이에서는 큰 차이를 나타내지 않았으나 1.7 kb promoter 형질전환생쥐의 경우 유선 이외 조직에서도 발현되는 양상을 나타내었고, 3.1kb promoter line에서는 유선특이적으로 발현시키는 양상을 나타내었다. 그러므로 bovine $\beta$-casein promoter의 1.7 kb와 3.1 kb 사이에 유선특이적 발현을 유도하는 조절부위가 있을 것으로 추정된다.

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DNA fingerprinting patterns of 5 serotypes of Actinobacillus actinomycetemcomitans (Actinobacillus actinomycetemcomitans의 혈청형별 제한절편장 다변화에 관한 연구)

  • Choi, Jeom-Il;Koh, Myung-Yun;Yun, Il
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.365-375
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    • 1996
  • 5 serotypes(a, b, c, d, e) of Actinobacillus actinomycetemcomitans showed distinct hybridization patterns(DNA fingerprinting patterns) when the bacterial DNA were hybridized with randomly cloned 4.7-Kb sized DNA probe. The sizes of hybridized bands in each serotypes were different among serotypes and represented unique patterns of hybridization with the probe used. The serotype a showed two bands of fingerprinting patterns: 23.1 kb and 2.5 kb respectively. Serotype b and c showed single band: 6.6 kb and 9.5 kb, respectively. Serotype d and e showed two bands of hybridization: 23.1 kb and 2.8 kb, and 23.7 kb and 2.1 kb, respectively. The results indicate that this standard fingerpriting patterns of DNA hybridization with 4.7 kb probe can be further used for genotyping clinical isolates of Actinobacillus 8ctinomycetemcomitansand its relevance with periodontal disease activity.

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Molecular Cloning of nifH, D from Frankia EuIK1 Strain, A Symbiont of Elaeagnus umbellata Root Nodules (보리수나무 뿌리혹 공생균주인 Frankia EuIK1의 nifH, D클로닝)

  • Kim, Ho-Bang;Kim, Chun-Ho;Song, Seun-Dal;An, Chung-Sun
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.258-263
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    • 1994
  • Genomic Southern hybridization of Frankia EuIKl strain, a nitrogen fixing symbiont of Elaeagnus umbellate root nodules, with nifH,D of K. pneumoniae as a probe, showed that 3.2 Kb and 5.5 Kb of BamHI fragments and 15 Kb PstI fragment were strongly hybridized with the probe, indicating nifH,D are located on these fragments. Using the same probe, one clone(pEuNIF) was isolated from the genomic library constructed into pWE15 cosmid vector by colony hybridization. The 3.2 Kb and 5.5 Kb BamHI fragments of this clone were hybridized with the same probe and this result corresponds to the genomic Southern hybridization data. However, using nifH of Frankia FaCl strain as a probe, only the 3.2 Kb BamHI fragment showed hybridization signal. Amino acid sequence deduced from nucleotide sequence of 3' terminus of the 3.2 Kb and 5' terminus of the 5.5 Kb fragments showed that the former was highly homologous with that of ArI3 nifD from 182nd to 240th amino acids, while the latter was from 241st to 282nd amino acids. These results show that nifH and partial nifD sequences are located on the 3.2 Kb fragment and residual sequences of nifH on the 5.5 Kb fragment which is contiguous to the 3.2 Kb fragment.

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