A novel, single plasmid, approach to in-frame fusion.
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.
Orum
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Genetic Engineering Group, Denmark Technical University, Lyngby.
Poulsen
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LK
D004247:DNA / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D010957:Plasmids; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D004247:DNA; C044171:endodeoxyribonuclease BSSHII; D015252:Deoxyribonucleases, Type II Site-Specific