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Nucleic Acids Research, 1989, Vol. 17, No. 8 3107-3115
© 1989

MOLECULAR BIOLOGY

A novel, single plasmid, approach to in-frame fusion

Henrik Ørum*,+ and Lars K. Poulsen

The Genetic Engineering Group. Denmark Technical University Building 227, DK-2800 Lyngby, Denmark

*To whom correspondence should be addressed

Received February 17, 1989. Accepted March 15, 1989.

The sequence GCGCGCGCGC contains three overlapping, mutually exclusive, BssHII restriction sites, each corresponding to one of the three different reading frames. When inserted into a plasmid and digested with BssHII, the three sites in this sequence are cleaved at an approximate ratio of 2:1:2. consequently, this system can be used to simplify in vitro in frame fusions to involve a single plasmid. We have constructed such a plasmid and used it to select an open reading frame in a 1.6 kb cDNA fragment.

+Present address: Panum Institute, University of Copenhagen, Department of Biochemistry B, Blegdamsvej 3c, DK-2200 Copenhagen N., Denmark


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