• 제목/요약/키워드: Lambda Red recombination

검색결과 6건 처리시간 0.019초

A Series of Vectors with Alternative Antibiotic Resistance Markers for Use in Lambda Red Recombination

  • Quick, Laura N.;Shah, Ashka;Wilson, James W.
    • Journal of Microbiology and Biotechnology
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    • 제20권4호
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    • pp.666-669
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    • 2010
  • A target bacterial strain of interest for use in Red-based recombineering may already encode resistance to antibiotic markers used with current Red recombination tools, such that the resistance cannot be removed. Such cases include those where markers are needed to maintain an unstable genetic element co-resident in the strain or those where the genetic source of resistance is not known. We report the availability of PCR templates with FRT-flanked mutagenesis cassettes and plasmids encoding Red recombination functions that contain marker combinations not currently available on widely disseminated lambda Red molecular reagents. The functionality of these convenient alternative tools is demonstrated.

A Modified PCR-Directed Gene Replacements Method Using $lambda$-Red Recombination Functions in Escherichia coli

  • KIM SANG-YOON;CHO JAE-YONG
    • Journal of Microbiology and Biotechnology
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    • 제15권6호
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    • pp.1346-1352
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    • 2005
  • We have developed a modified gene replacement method using PCR products containing short homologous sequences of 40- to 50-nt. The method required $\lambda$-Red recombination functions provided under the control of a temperature-sensitive CI857 repressor expressed from the $P_{lac}$ promoter in the presence of IPTG on an easily curable helper plasmid. The method promoted the targeted gene replacements in the Escherichia coli chromosome after shifting cultures of the recombinogenic host, which carries the helper plasmid, to $42^{\circ}C$ for 15 min. Since this method employs $\lambda$-Red recombination functions expressed from the easily curable helper plasmid, multiple rounds of gene replacements in the E. coli chromosome would be possible. The procedures described herein are expected to be widely used for metabolic engineering of E. coli and other bacteria.

LED용Mg2+·Ba2+Co-Doped Sr2SiO4:Eu 노란색 형광체의 발광특성 (Luminescence Characteristics of Mg2+·Ba2+ Co-Doped Sr2SiO4:Eu Yellow Phosphor for Light Emitting Diodes)

  • 최경재;지순덕;김창해;이상혁;김호건
    • 한국세라믹학회지
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    • 제44권3호
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    • pp.147-151
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    • 2007
  • An improvement for the efficiency of the $Sr_{2}SiO_{4}:Eu$ yellow phosphor under the $450{\sim}470\;nm$ excitation range have been achieved by adding the co-doping element ($Mg^{2+}\;and\;Ba^{2+}$) in the host. White LEDs were fabricated through an integration of an blue (InGaN) chip (${\lambda}_{cm}=450\;nm$) and a blend of two phosphors ($Mg^{2+},\;Ba^{2+}\;co-doped\;Sr_{2}SiO_{4}:Eu$ yellow phosphor+CaS:Eu red phosphor) in a single package. The InGaN-based two phosphor blends ($Mg^{2+},\;Ba^{2+}\;co-doped\;Sr_{2}SiO_{4}:Eu$ yellow phosphor+CaS:Eu red phosphor) LEDs showed three bands at 450 nm, 550 nm and 640 nm, respectively. The 450 nm emission band was due to a radiative recombination from an InGaN active layer. This 450 nm emission was used as an optical transition of the $Mg^{2+},\;Ba^{2+}\;co-doped\;Sr_{2}SiO_{4}:Eu$ yellow phosphor+CaS:Eu red phosphor. As a consequence of a preparation of white LEDs using the $Mg^{2+},\;Ba^{2+}\;co-doped\;Sr_{2}SiO_{4}:Eu$ yellow phosphor+CaS:Eu red phosphor yellow phosphor and CaS:Eu red phosphor, the highest luminescence efficiency was obtained at the 0.03 mol $Ba^{2+}$ concentration. At this time, the white LEDs showed the CCT (5300 K), CRI (89.9) and luminous efficacy (17.34 lm/W).

Biosynthesis of Polymyxins B, E, and P Using Genetically Engineered Polymyxin Synthetases in the Surrogate Host Bacillus subtilis

  • Kim, Se-Yu;Park, Soo-Young;Choi, Soo-Keun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제25권7호
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    • pp.1015-1025
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    • 2015
  • The development of diverse polymyxin derivatives is needed to solve the toxicity and resistance problems of polymyxins. However, no platform has generated polymyxin derivatives by genetically engineering a polymyxin synthetase, which is a nonribosomal peptide synthetase. In this study, we present a two-step approach for the construction of engineered polymyxin synthetases by substituting the adenylation (A) domains of polymyxin A synthetase, which is encoded by the pmxABCDE gene cluster of Paenibacillus polymyxa E681. First, the seventh L-threonine-specific A-domain region in pmxA was substituted with the L-leucine-specific A-domain region obtained from P. polymyxa ATCC21830 to make polymyxin E synthetase, and then the sixth D-leucine-specific A-domain region (A6-D-Leu-domain) was substituted with the D-phenylalanine-specific A-domain region (A6-D-Phe-domain) obtained from P. polymyxa F4 to make polymyxin B synthetase. This step was performed in Escherichia coli on a pmxA-containing fosmid, using the lambda Red recombination system and the sacB gene as a counter-selectable marker. Next, the modified pmxA gene was fused to pmxBCDE on the chromosome of Bacillus subtilis BSK4dA, and the resulting recombinant strains BSK4-PB and BSK4-PE were confirmed to produce polymyxins B and E, respectively. We also succeeded in constructing the B. subtilis BSK4-PP strain, which produces polymyxin P, by singly substituting the A6-D-Leu-domain with the A6-D-Phe-domain. This is the first report in which polymyxin derivatives were generated by genetically engineering polymyxin synthetases. The two recombinant B. subtilis strains will be useful for improving the commercial production of polymyxins B and E, and they will facilitate the generation of novel polymyxin derivatives.

염료폐수 분해를 위한 가시광 감응형 Pt-C-TiO2 광촉매의 합성 (Synthesis of Visible-working Pt-C-TiO2 Photocatalyst for the Degradation of Dye Wastewater)

  • 한미선;윤창연;이종협
    • 청정기술
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    • 제11권3호
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    • pp.123-128
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    • 2005
  • $TiO_2$는 금속 산화물의 일종으로서 자체가 가지고 있는 물리화학적 안정성, 무독성, 탁월한 유기물의 산화분해력 등으로 인해 저농도의 환경 유해물질 정화 분야로 응용이 활발히 연구되고 있는 반도체 물질이다. 그러나 $TiO_2$는 자외선 영역대(${\lambda}$ < 387 nm, 태양광의 2.7%가 UV)의 빛을 통해서 활성을 나타내고, 여기된 전자의 빠른 전자-정공 재결합속도로 인해 광 효율이 저하되는 단점을 갖는다. 따라서 광 감응 파장대를 넓히고 재결합속도를 길게 함으로써 광효율을 높이고, 광촉매 활성을 증대하는 방향으로 연구의 초점이 모아지고 있는 실정이다. 본 연구에서는 $TiO_2$ 광촉매의 광 감응 파장대를 가시광선 영역으로 확대함과 동시에 여기된 전자와 정공의 재결합시간을 연장하기 위하여 백금(Pt)이 광침적(photodeposition)된 탄소(C) 도핑 $TiO_2$를 제조하였다. 제조한 $Pt-C-TiO_2$의 특성은 전자투과현미경(Transmission Electron Microscopic; TEM), 질소흡탈착법(Brunauer-Emmett-Teller method; BET), X-ray 회절 분석법(X-ray Diffractometer; XRD), 분광 산란 광도계(UV-visible diffuse reflectance spectroscopy; UV-Vis DRS), X-ray 광전자 광도계(X-ray Photoelectron Spectroscopy; XPS)를 통하여 살펴보았다. $Pt-C-TiO_2$의 광촉매 활성을 검증하기 위하여 아조 계열의 붉은색 염료인 Acid Red 44 ($C_{10}H_7N=NC_{10}H_3(SO_3Na)_2OH$)의 광분해 실험을 수행하였다. 광원은 Xe arc 램프(300 W, Oriel)를 사용하였으며 420 nm 이하 제거 필터를 사용하여 가시광 영역대의 빛만을 조사되도록 하였다. 그 결과, 제조한 $Pt-C-TiO_2$는 가시광선 하에서 사용제품과 비교하여 월등히 뛰어난 분해력을 보이며 $C-TiO_2$의 활성을 한 층 더 향상시킴을 확인하였다. 이는 무한 에너지 자원인 태양광을 이용한 염료 폐수 정화 시스템 응용으로의 유용한 결과라 할 수 있겠다.

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