For the PET Project at the InnovaBio lab, the goal was to grow a linked protein of PETase and MHETase in Brewer's Yeast. This was because the result would be the breakdown of Polyethylene Terephthalate (or plastic) and have the waste product of ethanol that could be used in a variety of ways. However, when the yeast was grown, no protein bands of any kind appeared on an SDS-PAGE gel or Western Blot after the yeast were lysed and sonicated. With the expectation of seeing the protein expressed after being lysed and sonicated, the project made a transition from yeast to Escherichia coli with the same pYES2 plasmid as it was compatible and could also be transformed into E. coli. Here, I present the pYES2 plasmid in BL21 E. coli. SDS-PAGE gels of the induced E. coli show that we were successful at lysing and sonicating but lack the expression of the protein. Double digest and PCR of purified plasmid show that the protein is present in the E. coli. These findings show that the sequence was successfully inserted into the pYES2 plasmid. However, further testing is needed to verify sequencing of the protein as well as to determine why galactose is not causing induction.
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