Team:USP-Brazil/Results:Assemblies

Results

Assemblies

To explore the questions on BioBricks characterization, we planned the construction of the following strains:

Figure 1: Map of planned test strains. DNA construction images hiding RBS and transcription stop sequences.

The P_AOX1 strains are basically different combinations of variants of only three DNA elements: the P_AOX1 promoter, the RFP reporter protein and the modified Mxr1p transcription factor (aforementioned on Detecthol {link aqui}). Thanks to Life Technologies, the strain B1 and the Mxr1p modified and minimum were synthetized and this enabled the possibility of construction of all other strains.

Comparing the fluorescence time delay and intensity between strains Control 1, A1, and B1, we will be able to check the strength and the response velocity of the device with modified P_AOX1 and the efficiency of codon-optimization on RFP for P. pastoris. The comparison between control strains 2 and 3 with A2 and B2 will draw the picture of PAOX1 behavior with the modified Mxr1p transcription factor with the same variables from the last comparison, but also testing if the shorter version of Mxr1p—consisting in the sequence for the N-terminal 400 aminoacids from Mxr1p—cited on literature [2] will work properly.

Figure 2: {Precisamos de uma imagem que mostre a diferença da sequência entre o Paox modificado e o normal e o Mxr1p modificado e o mínino com o normal – aquela costumeira imagem de uma linha representando a dupla fita e linhas abaixo dela representando as outras duplas-fitas em comparação com a anterior}