By E. Magnien (auth.), E. Magnien (eds.)
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Extra info for Biomolecular Engineering in the European Community: Achievements of the Research Programme (1982 – 1986) — Final Report
Such enzymes would be obvious assets to the S. carnosus starter cultures and, accordingly, the r(-amylase gene of B. stearothermophUus has been introduced in S. ~rnosus by F. Gatz (MUnchen). The work involved eetting up a cloning system for S. carnosus : DNA transfer was achieved by polyethylene glycol induced protoplast transformation, with an efficiency comparable to that with eaCh treated E. coli cells. Vectors were constructed by associating a S. aureus derived replicon to various antibiotic determinants and these were used for cloning the Ol-amylase gene.
Expression was poor (lX of that of the parental Bacillus strain) and the recombinant plasmid 51 was unstable. Instability was successfully dealt with by keeping to a minimum the amount of B. stearothermophilus DNA carried by the recombinant. Overexpression and enhanced (I-amylase secretion are presenUy being attempted, by fitting the coding sequence with the relevant regulatory signals from the S. carnosus lipase gene. 2. Yeasts. ~.! cerevisiae strains with new fermenting properties. lL~!!. , Kluyveromyc~.!
Both groups are currently exploring the expression of the protein as a fusion protein at the COOH terminus of B-galactosidase. Using such material McCrae et al. have been succesful in inducing low levels of neutral ising antibody. A useful byproduct of this project has been the production of cDNA clones of some of the other genes of these viruses and Cohen et al. have shown that they can be the basis of a diagnostic assay for the presence of virus that is more rapid and up to 10 times more sensitive than the currently available tests.