Team:Newcastle/16 July 2010

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{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
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=LacI BioBrick Construction=
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=Research=
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==Aims==
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We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.
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*To use PCR to extract ''lacI'' (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
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*The aim of today's experiment is to '''screen for the lacI insert'''. This could be done using PCR but that would require specific primers and is not as reliable.
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#Klein C, Kaletta C and Entian KD. (1993). "Biosynthesis of the lantibiotic subtilin is regulated by a histidine kinase/response regulator system". ''Applied and environmental microbiology''. 59(1). 296-303.  
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#Heinzmann S, Entian KD and Stein T. (2006). "Engineering Bacillus subtilis ATCC 6633 for improved production of the lantibiotic subtilin". ''Applied microbiology and biotechnology''. 69(5). 532-6.  
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== Protocol ==
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#Miniprep - the [[Team:Newcastle/Qiagen_Minipreps|miniprep]] protocol was followed.  
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#Restriction digests - the [[Team:Newcastle/Restriction_digests|restriction digests]] protocol was followed to allow the length of the DNA to be checked using single and double digest.  
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#*Single digest with Pst1
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#*double digests with Pst1 and Xba1
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#Gel electrophoresis - the protocol for [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] was followed.  
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#Set up liquid broth culture in LB. The protocol for [[Team:Newcastle/Growing_an_overnight_cultures| growing an overnight culture]] was followed.
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==Results==
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*The single digests (lanes 1-7) and the double digests (lanes 9-15) are separated by the molecular marker (lane 8).
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[[Image:Newcastle LacI BioBrick Construction Gel.png|700px|centre]]
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Figure 1: Gel electrophoresis of the single and double digests using Pst1 and Xba1.
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Inference
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{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 20:28, 25 October 2010

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Research

We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.

  1. Klein C, Kaletta C and Entian KD. (1993). "Biosynthesis of the lantibiotic subtilin is regulated by a histidine kinase/response regulator system". Applied and environmental microbiology. 59(1). 296-303.
  2. Heinzmann S, Entian KD and Stein T. (2006). "Engineering Bacillus subtilis ATCC 6633 for improved production of the lantibiotic subtilin". Applied microbiology and biotechnology. 69(5). 532-6.
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