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Team:Trieste/notebook
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The Jolly JoCare
a safe probiotic platform for protein expression
Week 1
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Suicide System
We prepared the antibiotics stock ( Amp, Kan, Cm, Spec ) and the DH5-alpha competent cells. We extracted the following BB from the kit: the constitutive promoter (BBa_J23100), the strong RBS (BBa_B0034), the weak RBS (BBa_B0031), the GFP (BBa_E0240) the double terminator (BBa_B0015), the toxin Tse2 (BBa_K314200). E.coli DH5-alpha competent cells were transformed with 2 ul of each BB extracted. We learnt the safety rules and mostly of the lab techniques.Antibody
The first week in the lab we prepared competent E. coli strains DH5L, W3110 and HB2151 necessary for future experiments. Competent W3110 and HB2151were first transformed with p-REP 4 plasmid which codes for Lac repressor. The recombinant colonies selected on plate containing kanamycin were inoculated the and made them competent. We also digested and purified the synthetized genes from pUC57 vector.Chassis
We received the two plasmids that we had built and ordered. The first one contains the RBS - CymR - SV5 tag (730bp), the second one contain T5 promoter - Cumate operator (129 bp).
