Team:LMU-Munich/Weekly Journal

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Revision as of 11:39, 4 September 2012

The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".

[ more news ]

Weekly Journal

Legend:

+--+--+--+--+--+--	+--+--+--+--+--	+--+--+--+--+--+--	+--+--+--+--+--+--
Bacillus Intro	Bacillus BioBrickBOX	SporeCoat FusionProteins	Germination STOP

	3-7 September 2012
	Plates of our spores diluted at 10^-2, 10^-4 and 10^-6 from the germination assay show NO GERMINATION for our triple and quadruple mutants, and plenty of germination for the WT positive control! We will try plating undiluted mutant spores to see if any germination occurs.
	26-31 August 2012
	Finally, we got our first glowing spore!! After 4 months of hard work we have our first proof that our system works.
	Jara created quadruple mutants using two variations on past mutants: cwlD::kan + cwlJ::spec + gerD::cat + sleB::mls and gerD::cat + sleB::mls + cwlJ::spec + cwlD::kan. Germination assay performed on triple and quadruple mutants.
	20-24 August 2012
	Double negative loop with lacZa finished -> works qualitatively (Julia)
	13-17 August 2012 Jara and Jenny used last week's double mutants to create triple mutants as follows: cwlD::kan + sleB::mls + cwlJ::spec ; cwlD::kan + sleB::mls + gerD::cat ; cwlD::kan + cwlJ::spec + gerD::cat ; gerD::cat + sleB::mls + cwlJ::spec.
	6-10 August 2012
	Jara and Jenny created four double-mutants using resistance-cassettes to knock out germination genes as follows: cwlD::kan + sleB::mls ; gerD::cat + sleB::mls ; gerD::cat + cwlD::kan ; cwlJ::spec + cwlD::kan. We also created the resistance cassette knockout cwlB::kan.
	30 July - 3 August 2012

	23-27 July 2012
	Plate reader measurements of the Bacillus promoters P_liaG, P_liaI and P_lepA finished. Look at Data!
	16-20 July 2012

Plate reader measurements of the Anderson promoters J23100, J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118 in the Bacillus reporter vector pSB_Bs3C-luxABCDE completed. Look at Data!

Cloning of the Anderson promoters J23100, J23102, J23103, J23106 in the reporter vector pSB_Bs1C-lacZ for β-galactosidase assays finished.

9-13 July 2012

2-6 July 2012

Clean deletions of germination genes sleB and cwlB from PCR accomplished. DNA purified and frozen to be later transformed with Bacillus.

β-galactosidase assays of the promoters P_liaG, P_liaI and P_veg are finished. Look at Data!

Cloning of the Anderson promoters J23100, J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118 in pSB1C3 finished.

25-29 June 2012

18-22 June 2012

11-15 June 2012

Clean deletions of germination genes cwlD, cwlJ, and gerD from PCR accomplished. DNA purified and frozen to be later transformed with Bacillus.

4-8 June 2012

28-1 June 2012

21-25 May 2012

Promoters P_liaG, P_liaI, P_veg and P_lepA are now in the vector pSB1C3 as BioBrick standard for the registry. BioBricks are BBa_K823000, BBa_K823001, BBa_K823002, BBa_K823003.

The Anderson promoters J23100, J23101, J23102, J23103, J23106, J23107, J23113, J23114, J23115, J23117, J23118 are now in the Bacillus reporter vector pSB_Bs3C-luxABCDE for measuring their activity as luminescence.

The Bacillus promoters P_liaG, P_liaI and P_lepA are in the Bacillus reporter vector pSB_Bs3C-luxABCDE.

14-18 May 2012

7-11 May 2012

Cloning of P_liaG, P_liaI and P_veg in vector pSB_Bs1C-lacZ finished.

30 April-4 Mai 2012

23-27 April 2012

16-20 April 2012

9-13 April 2012

Jara created the single mutant cwlD::kan.

2-6 April 2012

Jara successfully created our first single knockouts of germination genes using a resistance cassettes: sleB::mls, gerD::cat and cwlJ::spec.

Jara tried knocking out cwlB using the kan resistance cassette. Mutants of cwlB::kan grew very poorly.

26-30 March 2012

Jara tried knocking out cwlB using the tet resistance cassette. Mutants of cwlB::tet grew very poorly.

19-23 March 2012

We decided which genes to knock out for the germination stop. Based on the work of [http://www.ncbi.nlm.nih.gov/pubmed/19554258 J. Kim and W. Schumann (2009)], we decided to knock out genes cwlB, gerD, cwlJ, and sleB. From the research of [http://www.ncbi.nlm.nih.gov/pubmed/11466293 B. Setlow et al (2001)], we also chose cwlD.

12-16 March 2012

5-9 March 2012

27 February - 2 March 2012

20-24 February 2012

Team fully formed!

Antibiotic abbreviation legend:

cat: chloramphenicol

kan: kanamycin

mls: Macrolide-Lincosamide-Streptogramin B

spec: spectinomycin

tet: tetracycline

Retrieved from "http://2012.igem.org/Team:LMU-Munich/Weekly_Journal"

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+|Plates of our spores diluted at 10^-2, 10^-4 and 10^-6 from the germination assay show NO GERMINATION for our triple and quadruple mutants, and plenty of germination for the WT positive control! We will try plating undiluted mutant spores to see if any germination occurs.
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 <img src="https://static.igem.org/mediawiki/2012/f/f6/GerminationSTOP.png" height=40"/></a></html>
 |Jara created quadruple mutants using two variations on past mutants: ''cwlD''::kan + ''cwlJ''::spec + ''gerD''::cat + ''sleB''::mls and ''gerD''::cat + ''sleB''::mls + ''cwlJ''::spec + ''cwlD''::kan.
+Germination assay performed on triple and quadruple mutants.
+|
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 <html><a href="https://2012.igem.org/Team:LMU-Munich/Germination_Stop">
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-Jenny presented ''Bacillus'' germination review to the group. We discussed stages of germination & how our mutants would prevent germination from occurring. Discussed possible germination causes besides nutrients, and how to hopefully prevent them. We realized partial germination caused by biochemical reactions from nutrients might cause the spores to deform, even if they did not complete germination. Therefore Jenny began research on this possibility.
 Clean deletions of germination genes ''sleB'' and ''cwlB'' from PCR accomplished. DNA purified and frozen to be later transformed with ''Bacillus''.