Production of eukaryotic proteins and nucleic acid molecules in c. elegans

摘要

Plasmid vectors for expression in Caenorhabditis elegans harbouring a heat inducible promoter nucleotide sequence, a synthetic intron nucleotide sequence optionally containing a Shine-Dalgarno sequence for efficient shuttling between C. elegans and E. coli, optionally a nucleotide sequence coding for a nuclear localisation signal or secretion signal, a nucleotide sequence coding for a recognisable tag, optionally a nucleotide sequence coding for a fluorescent protein, a nucleotide sequence coding for a protease cleavage site, a multiple cloning site containing a nucleotide sequence coding for an eukaryotic, such as human, protein or a nucleic acid molecule and a nucleotide sequence coding for termination of translation, are described. Methods of particularly large scale production of eukaryotic, such as human, proteins and nucleic acid molecules in nematodes are also described. The methods comprise injection one or several different plasmid vectors into the gonad of C. elegans hermaphrodites, cultivating the nematodes in growth medium, optionally comprising labeled bacteria as feed for the nematodes, at temperatures below 25°C, followed by shifting to 30 -33°C for induction of eukaryotic protein or nucleic acid molecule expression in several hundred somatic cells, with highest expression levels in neuronal and epidermal cells, and isolating the eukaryotic protein or nucleic acid molecule from said cells.