Latest revision as of 20:19, 25 October 2010

Research

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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+=Research=
-=LacI BioBrick Construction=
+We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.
-==Aims==
+#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.
-*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).
+#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.
-*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.
-#Miniprep - the [[Team:Newcastle/Qiagen_Minipreps|miniprep]] protocol was followed.
-#Restriction digests - the [[Team:Newcastle/Restriction_digests| restriction digest]] protocol was followed.
-#*Single digest with Pst1
-#*double digests with Pst1 and Xba1
-#Gel electrophoresis
-Set up liquid broth culture in 5 ml of LB - 1 colony from colonies 1-7
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