• 제목/요약/키워드: Pvu II

검색결과 49건 처리시간 0.011초

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

  • 김희정;이강민
    • KSBB Journal
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    • 제9권1호
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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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Bacillus thuringiensis serovar israelensis 4Q1로부터 분리된 plasmid 제한효소지도 작성 (Restriction endonuclease maps of three plasmids from bacillus thuringiensis serovar israelensis 4Q1)

  • ;이영근;강석권
    • 미생물학회지
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    • 제23권2호
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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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Streptomyces coelicolor A3(2)에서 hrdA유사 Sigma 인자 유전자의 클로닝 (Cloning of hadA-like Sigma Factor Gene from Streptomyces coelicolor A3(2))

  • 한지숙;조은정;노정혜
    • 미생물학회지
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    • 제32권4호
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    • pp.264-270
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    • 1994
  • 세균의 RNA 중합효소에서 여러 ${\sigma}$ 인자들 간에 보존된 아미노산 서열중 2.3 부위와 4.2 부위의 아미노산 서열로부터 유 n하여 두가지의 PCR primer를 제작하였다. 이들을 이용하여 PCR을 수행하였을 때, E. coli와 Streptomyces coelicolor의 DNA로부터 예상되었던 480 bp 정도의 DNA가 증폭되는 것을 관찰하였다. E. coli DNA에서 증폭된 DNA를 클로닝하여 염기서열을 결정한 결과 E. coli의 rpoS 유전자로부터 유래하였음을 알았다. 이를 탐침으로 S. coelicolor에서 genomic DNA hybridization을 수행하였을 때, PvuII 절편 두가지 (3.5 kb, 2.0 kb) 와 SalI 절편 두가지(3.4kb, 1.5 kb)에 탐침이 결합하는 것을 관찰하였다. 3.5 kb의 pvuII 절편을 sublibrary로부터 클로닝하고, 탐침이 결합하는 1.0kb의 BamHI/HincII 절편의 염기서열을 분석하였다. 부분적으로 결정된 염기서열을 BLAST 프로그램을 이용하여 GenBank와 EMBL, PDB 등의 data library의 유전자들과 비교하여 본 결과Streptomyces속의 ${\sigma}$인자들을 비롯한 Synechococcus종, Anabaena종, Pseudomonas aeruginosa, Stigmatella aurantica 등의 주된 ${\sigma}$ 인자와 높은 유사성을 보였다. 현재까지 1.2 부위와 4 부위에 해당하는 부분의 염기서열을 결정하였는데, 이 부분은 S. coelicolor에서 알려진 다섯가지의 ${\sigma}$ 인자 유전자 중 hrdA와 가장 높은 유사성을 보이며, 아미노산의 유사성이 1.2부위에서는 88%, 4 부위에서는 75%인 것으로 나타났다.

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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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    • 제50권5호
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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
    • 한국환경성돌연변이발암원학회지
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    • 제22권1호
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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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    • 제18권3호
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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.

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

  • 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.
    • 한국수정란이식학회지
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    • 제18권3호
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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.

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

  • 권용태;전희숙;노현모
    • 미생물학회지
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    • 제26권1호
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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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Yorkshire종 돼지에서 PCR-RFLP을 이용한 Estrogen Receptor의 유전적 다형과 산자수간의 관련성 (Association of Genetic Polymorphisms of Estrogen Receptor with Litter Size using PCR-RFLP in Yorkshire Swine)

  • 김지은;송원철;최봉도;고용;박성수;홍기창
    • Journal of Animal Science and Technology
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    • 제45권4호
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    • pp.523-528
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    • 2003
  • 본 연구는 PvuII PCR-RFLP를 이용하여 Yorkshire종 돼지에서 Estrogen Receptor의 유전적 다형과 산자수간의 관련성을 분석하기 위하여 수행되었다. 무창 돈사에서 사육중인 242두의 종빈돈으로부터 혈액을 채취하여 PvuII PCR- RFLP로 ER 유전자형을 결정하였다. ER 유전자 좌위에서 유전자 빈도는 각각 0.39(A)와 0.61(B)였다. ER 유전자형별 산자수에 대한 효과는 최소제곱평균을 설명하는 고정모형을 설정하여 추정하였다. 분석결과 복당 총산자수와 실산자수에서 특정 ER 대립유전자(B)에서 산자수 증진효과가 관찰되었다. 따라서 돼지 ER 좌위의 유전적 변이는 번식돈의 산자수 증대와 관련된 marker-assisted selection(MAS)에 응용될 수 있을 것으로 사료된다.