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From 2012.igem.org

Labjournal

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Week 2 (05/07 - 05/13/12)

• Doing the first steps to prepare at the Student Acadamy in cooperation with the CeBiTec. Finding suitable BioBricks to develop an colurful and easy experiment. Trying to isolate the GFP and RFP containing plasmid and to transform an Plasmidmix to produce colored Agar-plates.

• Primerdesign for isolation of laccases from genomic DNA of Xanthomonas campestris B100, E. coli BL21(DE3) and Bacillus pumilus ATCC7061. The forward primers were designed with T7 promotor-overhanging ends after prefix and the first 20 bases of the wanted gene. The reverse primers were designed with a HIS-Tag and tweo stop codons before suffix and the last 20 bases of the wanted gene without the stop codon.

• Successful PCRs of laccase genes CopA from Xanthomonas campestris B100, CueO from E. coli BL21(DE3) with the isolated genomic DNA as template.

• Because we want to characterise laccases from different bacteria we had to order the bacterial strains, which weren't available at the University Bielefeld, from [http://www.dsmz.de/|DSMZ].

• [http://www.dsmz.de/catalogues/details/culture/DSM-7039.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Thermus thermophilus HB27]

• [http://www.dsmz.de/catalogues/details/culture/DSM-18197.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Bacillus halodurans C-125]

• [http://www.dsmz.de/catalogues/details/culture/DSM-40069.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Streptomyces lavendulae REN-7]

• [http://www.dsmz.de/catalogues/details/culture/DSM-40236.html?tx_dsmzresources_pi5%5BreturnPid%5D=304|Streptomyces griseus IFO 13350]

• After some empty agarose gels we finally isolated laccase gene CotA from Bacillus pumilus ATCC7061 as a PCR product. As template we used the plasmid we got from the Swiss working group.
• Team Modeling: Looking for suitable Software and enzymkinetics to model the degradation of our substartes with the different laccases. Finding the Michaelis-Menten kinetics and matlab.