PubMed · 15584182
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G McGee, E T Juengst. 1999. Genetic enhancement.. https://pubmed.ncbi.nlm.nih.gov/15584182/
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Methylotrophic yeast Pichia pastoris is convenient for the expression of eukaryotic foreign proteins owing to its potential for posttranslational modifications, protein folding, and facile culturing. In this work, human interleukin (hIL)-2 was successfully produced as a secreted fusion form in recombinant P. pastoris. By employing green fluorescent protein (GFP) as a monitoring fusion partner, clear identification of fusion protein expression and quantification of intracellular hIL-2 were possible even though there was no correlation between culture supernatant fluorescence and secreted hIL-2 owing to high media interference. Importantly, by the addition of casamino acids in basal medium, we were able to enhance threefold amount of secreted hIL-2, which was present both as a fusion and as a clipped fragment.
In their jointly written book, From Chance to Choice: Genetics and Justice, Allen Buchanan, Dan Brock, Norman Daniels and Daniel Wikler defend 'the development and deployment of genetic intervention technologies..', including genetic enhancements, against charges that they exacerbate injustice. The present paper examines some of their arguments. The first section shows that the authors confuse real societies with just societies. The second shows that without this confusion, their arguments reveal the enormous justice-impairing potential of deploying genetic enhancements in such societies as the United States.
Efficient secretion of human lysozyme from the yeast, Kluyveromyces lactis, was achieved by using more stable vectors in the order of S11 replication origin-containing episomal vector < full-length K. lactis plasmid pKD1-containing vector < centromeric vector < chromosome-integrated vectors. Cells containing a PGK (phosphoglycerate kinase) promoter-driven integration vector grown in non-selective rich medium achieved the highest level of secretion, approximately 100 microg lysozyme secretion ml(-1) culture: this level was approximately 10-fold higher than that achieved by episomal vectors. An additional copy of the protein disulfide isomerase gene further facilitated the secretion.