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Nucleic Acids Research, 1981, Vol. 9, No. 20 5187-5202
© 1981

MOLECULAR BIOLOGY

Expression of plant tumor-specific proteins in minicells of Escherichia coli: a fusion protein of lysopine dehydrogenase with chloramphenicol acetyltransferase

Joachim Schröder*, Annette Hillebrand*, Werner Klipp+ and Alfred Pühler+

*Max-Planck-Institut für Züchtungsforschung Abt. Schell, D-5000 Köln 30, GFR +Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld D-4800 Bielefeld 1, GFR

Received August 14, 1981. Fragment EcoRI 7 from Ti-plasmid pTi Ach5, a part of the T-DNA in octo-pine tumors, was cloned in both orientations into pACYC184 and expressed in E.coli minicells. The cells synthesized four proteins from four different coding regions on EcoRI 7. Two of the proteins (Mr 25.000 and 26.000) were expressed with promoters from the Ti-plasmid fragment, while transcription for the two other proteins (Mr 18.000 and 74.000) started with a promoter on pACYCi84. The M 18.000 protein represented a fusion product between chlor-amphenicol acetyltransferase (CAT) on pACYC184 and a part of lysopine dehydrogenase (LpDH), the enzyme synthesizing octopine and lysopine in plant tumor cells. The results suggest that E.coli minicells are a valuable system to study the proteins coded for by the T-region of Ti-plasmids.


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[Abstract] [PDF]


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