• 제목/요약/키워드: Bacteriophage P2

검색결과 78건 처리시간 0.025초

Bacillus subtilis var, 816 Bacteriophage의 분리와 일반적 성질에 관한 연구 (Studies on the Isolation and Characterization of Bacteriophage of Bacillus subilis var, S16)

  • 이주식
    • 미생물학회지
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    • 제16권2호
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    • pp.71-78
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    • 1978
  • Bacillus subtilis var. 816 was used for manufacturing fermented soybean which in turn is used as flavoring agent. Fermentation of soyebean or flat wheat was occasionally failed. It was elucidated that failure was due to the presence of bacteriophage. According to Hemphill and Whitely (1975), this bacteriphage might be belonged to the viulent phage group I as it is similar to SP82G, ${\phi}25$. In fact, the phenomena of the increase of moisture, disappenarance of mucin and existence of undersirable bacteria was attributed to the contamination of the above phage during the course of fermentation of soybean or flat wheat. Particularly disapparance of mucin was sufficiently correlated by the replication of the bacteriophage. The above phage can grow in the range of $30^{\circ}C\;to\;70^{\circ}C$. The optimum temperature was $40^{\circ}C{\sim}50^{\circ}C$. The optimum pH range was between pH 7.4 and pH 8.0. It is noticeable that staphylococci was replicating simultaneously with the phage. The head of 816 phage is hexagonal with a diameter of $10{\times}165{\sim}10{\times}240\;nm$. The end of the tail is enlaged. It has a size of 25 nm and this end areas are spreaded widely as fingers.

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Preincubation without attB DNA inhibits In Vitro Integrative Recombination of P 1 Mutant attP DNA of Bacteriophage Lambda

  • Yoo, Seung-Ku
    • Journal of Microbiology and Biotechnology
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    • 제5권3호
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    • pp.132-137
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    • 1995
  • The lambda integrase (lnt) is believed to bind to several arm and core sites of attP DNA in order to facilitate intasome formation. We have done systematic mutagenic analysis on all 5 arm sites and found that P1 is absolutely required for integration while P2 is not. We also found that all 3 P' arm sites(P'1, P'2, and P'3) are required for efficient integrative recombination. P'1, which is an important binding site for excision, also seems to be crucial for integration when preincubation of attP DNA with Int and IHF is performed before recombination. Preincubation assay revealed that preincubation with Int and IHF improved the efficiency of recombination of wild type attP DNA and demolished recombinations of P'1 mutant attP DNAs.

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SP816 박테리오파아지의 생리적 특성 (Physiological characterization of SP816 bacteriophage)

  • 이오형
    • 미생물학회지
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    • 제24권2호
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    • pp.161-167
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    • 1986
  • Some of the physiological properties of Sp816 bacteriophage of Bacillus subtilis SNU816 were characterized. It could form plaques on either B. subtilis SNU816 or B. natto 8102, but not on any other bacillus strains investrgated. Its plaque morphology was circular with a diameter of less than 1.0mm and had a narrow halo surrounding the clear center. Its latent period was 34-36 min and had a burst size of 547. It was most stable at pH 6.0, and rapidly inactivated at $60^{\circ}C$ with a initial deaty rate of -0.216 $min^{-1}$. Host range, thermal inactivation rate at $60^{\circ}C$, pH stability, and UV sensitivity revealed that SP816 was quite different from any other phages investigated together but seemed to be rather related to B. natto phages.

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An analysis of the arm-type site binding domain of bacteriophage .lambda. integrase

  • Cho, Eun-Hee
    • Journal of Microbiology
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    • 제33권2호
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    • pp.165-170
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    • 1995
  • The 356 amino acid long lambda integrase protein of bacteriophage .lambda. constains two autonomous DNA binding domains with distinct sequence specificities. The amino terminal domain of integrase is implicated to bind to the arm-type sequences and the carboxyl domain interacts with the coretype sequencess. As a first step to understand the molecular mechanism of the integrase-DNA interaction at the arm-type site, the int(am)94 gene carrying an amber mutation at the 94th codon of the int was cloned under the control of the P$\_$tac/ promoter and the lacI$\_$q/ gene. The Int(am)94 mutant protein of amino terminal 93 amino acid residues can be produced at high level from a suppressor free strain harboring the plasmid pInt(am)94. The arm-type binding activity of Int(am)94 were measured in vivo and in vitro. A comparison of the arm-type binding properties of the wild-type integrase and the truncated Int(am)94 mutant indicated that the truncated fragment containing 93 amino acid residues carry all the determinants for DNA binding at the arm-type sites.

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Purification of Filamentous Bacteriophage M13 by Expanded Bed Anion Exchange Chromatography

  • Tau Chuan;Chee Kin;Wen Siang;Beng Ti;Wan, Wan-Mohammad;Arbakariya
    • Journal of Microbiology
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    • 제42권3호
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    • pp.228-232
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    • 2004
  • In this paper, we investigated the development of a simplified and rapid primary capture step for the recovery of M13 bacteriophage from particulate-containing feedstock. M13 bacteriophage, carrying an insert, was propagated and subsequently purified by the application of both conventional multiple steps and expanded bed anion exchange chromatography. In the conventional method, precipitation was conducted with PEG/NaCl, and centrifugation was also performed. In the single step expanded bed anion exchange adsorption, UpFront FastLine$\_$TM/20 (20mm i.d.) from UpFront Chromatography was used as the contactor, while 54$m\ell$ (H$\_$o/=15cm) of STREAMLINE DEAE (p=1.2 g/㎤) from Amersham Pharmacia Biotechnology was used as the anion exchanger. The performance of the two methods were evaluated, analysed, and compared. It was demonstrated that the purification of the M13 bacteriophage, using expanded bed anion exchange adsorption, yielded the higher recovery percentage, at 82.86%. The conventional multiple step method yielded the lower recovery percentage, 36.07%. The generic application of this integrated technique has also been assessed.

P2 sir3-sized head에 packaging 되기 용이한 크기의 박테리오파지 P4 유도체 조성과 정성 연구 (Construction and characterization of the bacteriophage P4 derivatives whose genome size suitable for packaging into a P2sir3-sized head)

  • 김경진
    • 미생물학회지
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    • 제51권1호
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    • pp.1-6
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    • 2015
  • "P2 sir-관련 도움파지 비효율성"이라는 용어는 P2 sir 변이체가 그들의 위성 박테리오파지인 P4에 대해 도움파지 역할을 충분히 못하는 것을 가리키는 용어이다. 이 연구의 목적은 이러한 P2 sir-관련 도움파지 비효율성을 극복하는 요인들을 조사하는 것이다. 우선 P2의 cos 영역을 함유하는 P4 sid71 cosP2가 P2 sir3의 도움파지 비효율성을 극복하는지를 조사하였다. 그 결과 P4 sid71 cosP2는 P2 sir3의 도움파지 비효율성을 극복하지 못하였다. P2의 cos 영역 대신에 $P2_{sir}$-sized head에 packaging되는 DNA 크기가 도움파지 비효율성을 극복하는 중요한 요인으로 나타났다. 이 연구에서는, DNA 조작을 통해 packaging 되는 DNA 크기가 P4 ost1과 P4 ost2 사이가 되는 세 종류의 P4 유도체를 조성하였다. 그 중 하나인 P4 sid71 delRI::apr의 CsCl 균등밀도차실험을 거쳐 packaging되는 DNA의 크기를 확인하였다. P4 유도체들의 후손방출량 실험에 따르면 그들은 모두 P2sir3-관련 도움파지 비효율성을 극복하였다. $P2_{sir3}$-sized head에 잘 packaging되는 P4 유도체의 크기는 28-29 kb로 나타났다.

DNA 형태 적응을 거쳐 P2sir-관련 도움파지 비효율성을 극복하는 박테리오파지 P4 sid+ 유도체 정성 연구 (Characterization of the bacteriophage P4 sid+ derivative overcoming P2sir-associated helper inefficiency through DNA conformational adaptation)

  • 김경진
    • 미생물학회지
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    • 제52권1호
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    • pp.120-124
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    • 2016
  • P2-크기 머리에 packaging 될 특정 DAN 크기(28-29 kb long)와 박테리오파지 P4 유전자 sid의 변이가 "P2 sir-관련 도움파지 비효율성"을 극복할 수 있는 요소로 압축되었다. 유전자 sid의 변이 여부가 필수적인지를 확인하기 위해, 정상적인 sid 유전자를 가지며 $P2_{sir3}$-크기의 큰 머리에 packaging 될 DNA 크기가 28.5 kb되는 P4 delRI::kmr을 사용하여 실험하였다. P4 delRI::kmr이 P2 sir3 용원소에 대해 낮은 EOP를 보이므로, 이를 증가시키기 위해 P2 sir3 용원소를 숙주세포로 하여 파지 stock을 제조하였다. 이 과정에서 P4 delRI::kmr이 P2 sir3 용원소에 대해 적응하는 것을 관찰하고, CsCl 부양 균등밀도 편차실험과 분리된 DNA의 전기영동을 통해 그것이 packaging 될 머리 크기에 따른 DNA 형태 변화에 의한 적응이라는 것을 알아냈다. P2 sir3 용원소에 적응된 P4 delRI::kmr과 적응되지 않은 P4 delRI::kmr stock의 burst size 결정 실험은, sid 유전자 변이에 상관없이 packaging 될 DNA 크기에 의해 "P2 sir-관련 도움파지 비효율성"이 극복된다는 것을 보여주었다.

층상이중수산화물에 의한 인공지하수내의 박테리오파지 T7 제거 (Removal of Bacteriophage T7 from Artificial groundwater by Layered Double Hydroxide)

  • 박정안;이창구;강진규;김성배
    • 대한환경공학회지
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    • 제33권6호
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    • pp.426-431
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    • 2011
  • 본 연구의 목적은 마그네슘-철 층상이중수산화물(Mg-Fe LDH)을 이용하여 인공 지하수에서 바이러스를 제거하는 것이다. Mg-Fe LDH를 이용한 박테리오파지 T7의 제거를 관찰하기 위하여 다양한 실험조건에서 회분실험을 실시하였다. 실험 결과, Mg-Fe LDH에 의한 T7 제거는 빠른 반응으로써, 2~3시간 안에 평형에 도달하였다. Mg-Fe LDH의 T7 제거능은 $1.57{\times}10^8pfu/g$이었고, 제거율은 96%이었다. 또한, pH 6.2~9.1 범위에서 용액 pH가 T7 제거에 미치는 영향은 미미하였다. 음이온들($SO_4^{2-}$, $CO_3^{2-}$, $HPO_4^{2-}$)이 T7 제거에 미치는 영향은 중요하였는데, 이유는 이들 음이온들이 LDH상의 흡착지점에 T7과 경쟁하기 때문이다. 반면, 질산염($NO_3^-$)이 T7 제거에 미치는 영향은 미미하였다. 본 연구에 의하면, Mg-Fe LDH는 흡착제로써 수처리 과정에서 바이러스제거에 적용될 수 있을 것으로 판단된다.

Characterization and Genomic Analysis of Novel Bacteriophage ΦCS01 Targeting Cronobacter sakazakii

  • Kim, Gyeong-Hwuii;Kim, Jaegon;Kim, Ki-Hwan;Lee, Jin-Sun;Lee, Na-Gyeong;Lim, Tae-Hyun;Yoon, Sung-Sik
    • Journal of Microbiology and Biotechnology
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    • 제29권5호
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    • pp.696-703
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    • 2019
  • Cronobacter sakazakii is an opportunistic pathogen causing serious infections in neonates. In this study, a bacteriophage ${\Phi}CS01$, which infects C. sakazakii, was isolated from swine feces and its morphology, growth parameters, and genomic analysis were investigated. Transmission electron microscopy revealed that ${\Phi}CS01$ has a spherical head and is 65.74 nm in diameter with a 98.75 nm contracted tail, suggesting that it belongs to the family Myoviridae. The major viral proteins are approximately 71 kDa and 64 kDa in size. The latent period of ${\Phi}CS01$ was shown to be 60 min, and the burst size was 90.7 pfu (plaque-forming units)/infected cell. Bacteriophage ${\Phi}CS01$ was stable at $4-60^{\circ}C$ for 1 h and lost infectivity after 1 h of heating at $70^{\circ}C$. Infectivity remained unaffected at pH 4-9 for 2 h, while the bacteriophage was inactivated at pH <3 or >10. The double-stranded ${\Phi}CS01$ DNA genome consists of 48,195 base pairs, with 75 predicted open reading frames. Phylogenetic analysis is closely related to that of the previously reported C. sakazakii phage ESP2949-1. The newly isolated ${\Phi}CS01$ shows infectivity in the host bacterium C. sakazakii, indicating that it may be a promising alternative to antibacterial agents for the removal of C. sakazakii from powdered infant formulas.