Purification Of Wild Type And G242r Mcad

For purification, 10 of the non-G985 mutations were subcloned into the pTrc-MCAD vector to allow expression of the mutant protein in higher yields, and transformed into E. coli TG1 cells. For the G799A mutation, the ECoRI/HindIII fragment (position 689-1315) of the pG242R plasmid was ligated into the pTrc-MCAD plasmid, digested with the same enzymes.

The wild type and the mutant G242R have successfully been purified. The enzymes were purified from 6L of culture. The G242R mutant was co-overexpressed with the

Table 1. Purification of wild type and G242R MCAD.

Protein Total activity Specific Activity

Protein Total activity Specific Activity

Table 1. Purification of wild type and G242R MCAD.

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