• Title/Summary/Keyword: Pvu II

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Solvent Effect on Restriction Endonuclease : Alteration of Specificity of Restriction Endonuclease PvuII in Hydrophobic Solution (제한효소에 대한 용매의 영향 :소수성 용매에 의한 PvuII 특이성 변화)

  • 김희정;이강민
    • KSBB Journal
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    • v.9 no.1
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    • pp.63-71
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    • 1994
  • During the last decade enzyme reaction in organic solvent has been studied to show that specificity in buffer is different from that in organic solvent. The specificity of restriction enzyme was effected by various factors such as ionic strength, salt organic solvent and temperature. In this study, restriction enzyme PvuII which is used most frequently in genetic engineering and the substrate was vector pGEM3 whose sequence was already known were used. As a result the recognition sequence site was changed in the presence of organic solvents whose Log P are -1.5∼0. Their specificities were contrast with activities were contrasted. Specificities were not changed in organic solvent easily in inactivating enzyme. We think that the enzyme recognition site was not changed randomly but by preferential order. A recombinant vector which does not contain typical cleavage site CAG↓CTG was cleaved in 20% ethanol solution. This result might show that restriction enzyme could be used to cleave at unusual sites by changing the reaction conditions.

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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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Cloning of hadA-like Sigma Factor Gene from Streptomyces coelicolor A3(2) (Streptomyces coelicolor A3(2)에서 hrdA유사 Sigma 인자 유전자의 클로닝)

  • Hahn, Ji-Sook;Cho, Eun-Jung;Roe, Jung-Hye
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.264-270
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    • 1994
  • A gene coding for a novel putative $\sigma$ factor of RNA polymerase has been identified from Streptomyces coelicolor A3(2) using Escherichia coli rpoS gene fragment as a probe. The 486 bp rpoS gene fragment was amplified from E. coli genomic DNA by PCR with two synthetic oligonucleotides, the sequences of which were deduced from the amino acid sequences in the regions 2.3 and 4.2 conserved among various bacterial factors. When E. coli genomic DNA fragments were hybridized with cloned rpoS probe, only one band corresponding to rpoS gene (3.2 kb PvuII fragment or 2.3 kb KpnI fragment) was detected. In S. coelicolor, however, two bands were detected both in PvuII digested DNA and SalI digested DNA. 3.5 kb PvuII fragment which binds the rpoS gene probe was cloned (pMS1) from the sublibrary, and the nucleotide sequences of 1.0 kb BamH'/HincII subclone (pBH2) was partially determined. The nucleotide sequences revealed extensive similarity to other $\sigma$ factor genes of S. coelicolor (hrdA, hrdB, hrdC, hrdD), S. aureofaciens (hrdA, hrdB, hrdC, hrdD), Synechococcus species, Pseudomonas aeruginosa, Stigmatella aurantiaca, and Anabaena species. The nucleotide sequences in regions 1.2 and 4 were compared with the corresponding regions of 5 known ${\sigma}$ factor genes of S. coelicolor by multiple alignment. It turned out that the cloned gene is most closely related to hrdA showing 88% amino acid similarity in region 1.2 and 75% in region 4.

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Association of Estrogen Receptor Gene Polymorphism in Patients with Degenerative Lumbar Spondylolisthesise

  • Lee, Jung-Sub;Suh, Kuen-Tak;Kim, Jeung-Il;Lim, Jong-Min;Goh, Tae-Sik
    • Journal of Korean Neurosurgical Society
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    • v.50 no.5
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    • pp.415-419
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    • 2011
  • Objective : The purpose of this study was to investigate the possible association of estrogen receptor alpha ($ER{\alpha}$) gene polymorphisms in a cohort of degenerative spondylolisthesis (DS) patients. Methods : Accordingly, the authors examined the association between DS and $ER{\alpha}$ gene polymorphisms in 174 patients diagnosed with DS. The $Pvu$ $II$ and $Xba$ $I$ polymorphisms, bone mineral density at the lumbar spine and femoral neck, and biochemical markers were analyzed and compared in the 174 patients with DS and 214 patients with spinal stenosis (SS). Results : A comparison of genotype frequencies in DS and SS patients revealed a significant difference for the $Pvu$ $II$ polymorphism only ($p$=0.0452). No significant difference was found between these two groups with respect to the $Xba$ $I$ polymorphism, BMD or biochemical markers. No significant association was found between the$Pvu$ $II$ polymorphism of $ER{\alpha}$ and BMD, vertebral slip or biochemical markers in patients with DS. Conclusion : These results suggest that the $ER{\alpha}$ gene polymorphism using $Pvu$ $II$ restriction enzyme influences the prevalence of DS.

Two Genetic Polymorphisms of the Human Lipoprotein Lipase Gene in Korean Patients with Essential Hypertension

  • Kang, Byung-Yong;Lee, Kang-Oh;Kim, Ki-Tae;Bae, Joon-Seol;Ryu, Jae-Chun;Kim, Jae-Hyoun;Lim, Seok-Rhin;Lee, Chung-Choo
    • Environmental Mutagens and Carcinogens
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    • v.22 no.1
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    • pp.22-29
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    • 2002
  • Essential hypertension is considered to be caused by a complicated combination of genetic and environmental factors. Alterations of lipid metabolism in plasma have been reported to be related to an increased risk of essential hypertension. The purpose of this study was to investigate the relationship between two genetic polymorphisms (Pvu II and Hind III RELPs) of the human lipoprotein lipase (LPL) gene and essential hypertension in korean population. In our result the Pvu II RFLP of LPL gene was significantly associated with essential hypertension (P < 0.05). Therefore, we suggest that the Pvu II RFLP of LPL gene may be useful as a genetic marker for essential hypertension in Korean population.

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Relationship between Genetic Variants of Mitochondrial DNA and Growth Traits in Hanwoo Cattle

  • Jeon, G.J.;Chung, H.Y.;Choi, J.G.;Lee, M.S.;Lee, C.W.;Park, J.J.;Ha, J.M.;Lee, H.K.;Sung, H.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.3
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    • pp.301-307
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    • 2005
  • Genetic variants of Hanwoo mtDNA in the region of cytochrome oxidase subunit I, II and III complex were detected using restriction enzymes. PCR primers were designed based on the bovine mtDNA sequence, and 6 primer sets (Mt4, Mt5, Mt6, Mt7, Mt8 and Mt9) were used. A total of 20 restriction enzymes were used, and 6 restriction enzymes, which were Hinf I, Pvu II, Rsa I, Eco RI, Bgl II, and Msp I, showed genetic polymorphisms. Significant associations between genetic variants and weight traits were observed at WT15 (p<0.05) and WT18 (p<0.01) with Pvu II for Mt9, Bgl II for Mt6 and Rsa I for Mt8 segments in the region of cytochrome oxidase subunit complex. Significant associations were also observed at Mt9-Pvu II and Mt6-Bgl II segments for WT9 (p=0.01), WT12 (p=0.02), respectively. These results suggest that genetic variants of mtDNA in the region of cytochrome oxidase subunit complex may be candidate segments for improvement of animal growth as weight traits.

Effects of Mitochondrial DNA Polymorphism on Growth Traits of Hanwoo (mt DNA 다형이 한우 성장에 미치는 영향)

  • Jeon, G.J.;Chung, H.Y.;Choi, J.G.;Lee, M.S.;Chung, Y.H.;Lee, C.W.;Park, J.J.;Ha, J.M.;Lee, H.K.;Na, K.J.
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.227-235
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    • 2003
  • 한우의 mt DNA cytochrome oxidase subunit I, II, 및 III complex지역의 유전적 다형현상을 제한효소를 이용하여 검출하였다. PCR primer 6종에 대하여 20가지 제한효소를 처리하였으며, Pst I, Pvu II, Rsa I, Eco RI, Bgl II, and Msp I 제한효소를 사용하여 유전적 변이를 검출하였다. 검출된 변이체와 한우의 성장과의 관련성을 조사한 결과 cytochrome oxidase subunit III complex 지역의 유전염기서열을 근거로 제작한 primer Mt9 좌위에서 제한효소 PvuII를 이용한 절단형과 체중형질 인 WT15(P<0.05) 및 WT18(P<0.01)에서 고도의 유의성이 관찰되었다. 아울러 , Mt9-Pvu II(P=0.07), Mt6-Bgl II(P=0.05), and Mt8-Rsa I(P=0.05) 좌위 또한 WT9, WTl5, and WT15에서 각각 통계적 유의성이 관찰되었다. 따라서 본 결과는 cytochrome oxidase subunit III complex segments가 candidate gene으로서 기초적 유전정 보 제공은 물론 유전적 개량을 위해 사용될 수 있을 것으로 사료된다.

Characterization of BmaI endonuclease from bacillus macerans ATCC 8244 (Bacillus macerans의 BmaI endonuclease의 특성에 대한 연구)

  • 권용태;전희숙;노현모
    • Korean Journal of Microbiology
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    • v.26 no.1
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    • pp.1-5
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    • 1988
  • The esolation and characterization of a new type II restriction endounclease, BamI, from Bacellus macerans ATCC 8244 were described. BmaI endonuclease was partially purified by procedures of ammonium sulfate fractionation, DEAE-cellulose and phosphocellulose chromatographies. This enzume recognized one site on pBR322 DNA, two sites on Bluescribe DNA, three sites on $\lambda$DNA and no site on SV 40 DNA. The same cleavage patterns for vareius DNAs as PvuI indicated that BamI is an isoschisomer of PvuI whose recognition sequence is 5'-CGATCG-3'. The optimal pH for the BmaI endonuclease activity was about 7.0 and optimal NaCl concentration was about 100mM. Manganese ion could partially replace magnesium as a cofactor, but calcium could not at all.

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Association of Genetic Polymorphisms of Estrogen Receptor with Litter Size using PCR-RFLP in Yorkshire Swine (Yorkshire종 돼지에서 PCR-RFLP을 이용한 Estrogen Receptor의 유전적 다형과 산자수간의 관련성)

  • Kim, J.E.;Song, W.C.;Choi, B.D.;Kho, Y.;Park, S.S.;Hong, K.C.
    • Journal of Animal Science and Technology
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    • v.45 no.4
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    • pp.523-528
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    • 2003
  • This study was performed to investigate a possible association of the porcine estrogen receptor(ER) locus with the total number of born(TNB) and number of born alive(NBA) in Yorkshire pigs. Using DNAs extracted from 242 Yorkshire pigs, the ER genotype was determined by PvuII PCR-RFLP. The ER allele frequencies of two types of A and B were 0.39 and 0.61, respectively. The least squares means of the litter size by ER genotype was evaluated. The TNB and NBA were found to be associated with an specific ER allele. The genotype at the porcine ER locus has an application potential for marker-assisted selection for litter size in pigs.