Dear all One of my human membrane proteins have been described to interact with additional subunits for its activity. To obtain functional form in yeast (Saccharomyces), I can think of two approach of either cloning all the subunits under one promoter or reconstitute in vitro.
For the first option, what is the length of base pairs between the stop codon of one gene and the start of Kozak sequence for the next one? Is there any preference for the order so that only one subunit is His tagged? Second option will need multiple purification steps and some trials with protein ratios. Is this better? Thank you. Theresa
