Team:Newcastle/15 July 2010

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=Research=
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=LacI BioBrick Construction=
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We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.
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==Aims==
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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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*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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#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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*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. The experiment is being repeated from yesterday as there are extra bandings
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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 digest]] protocol was followed.  
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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 [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] protocol was followed.  
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Set up liquid broth culture in 5 ml of LB - 1 colony from colonies 1-7
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{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 20:19, 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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