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

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.
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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-=LacI BioBrick Construction=
-==Aims==
+=Research=
-*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).
-*The aim of today's experiment is to '''screen for the lacI insert'''. The experiments that were carried out [[Team:Newcastle/16_July_2010#Protocol|yesterday]] are being repeated today as there were extra and unexpected bands on the image of the gel.
-==Protocol==
+We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.
-#Miniprep - the [[Team:Newcastle/Qiagen_Minipreps|miniprep]] protocol was followed.
-#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.
+#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.
-#*Single digest with Pst1
+#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.
-#*double digests with Pst1 and Xba1
-#Gel electrophoresis - the [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] protocol was followed.
-#Set up liquid broth culture in 5 ml of LB - 1 colony from colonies 1-7. The protocol for [[Team:Newcastle/Growing_an_overnight_cultures| growing an overnight culture]] was followed.
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Team:Newcastle/15 July 2010

From 2010.igem.org

Latest revision as of 20:19, 25 October 2010

Research