Double Tagged Serratia Marcescens Nuclease
AssigneeAbClonal Science, Inc.
InventorsDapeng Sun, Aine Quimby, Zhenyu Zhu
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Abstract
Either wild type or mutant Serratia Marcescens Nuclease (“SMN”) is engineered to display a C-terminal Chitin Binding Domain (CBD-tag), followed, at the C-terminus side of the CBD-tag, by a poly-histidine tag (His-tag), where the His-tag is preferably a 6-mer and preferably is preceded by a Gly-Ser linker, thereby generating a recombinant SMN protein that retains dual affinity tags (a CBD-tag and a His-tag) at the C-terminus, to make it easily removed from a reaction solution following digestion of nucleic acids in the reaction mixture. It can also be used for binding SMN to a solid support for use in nucleic acid digestion in a sample contacted with the solid support.
Claims19
A process of using a wild type or mutant recombinant Serratia Marcescens Nuclease having a Chitin Binding Domain tag at its C-terminus and with a poly-histidine tag on the C-terminal side of the CBD-tag, to digest nucleic acid in a reaction solution, and then removing it from the reaction solution, comprising:
- placing the recombinant Serratia Marcescens Nuclease in a reaction including nucleic acid for digestion under conditions for digestion to proceed;
- and
- removing the recombinant Serratia Marcescens Nuclease the from the reaction mixture by binding it to Chitin which is bound to a solid support and/or by binding it to metal ions.
A solid support bound to Chitin which is bound to a Chitin Binding Domain of a mutant recombinant Serratia Marcescens Nuclease having a Chitin Binding Domain tag at its C-terminus and with a poly-histidine tag on the C-terminal side of the CBD-tag.
A metal-ion containing solid support bound to a poly-histidine tag which is part of a mutant recombinant Serratia Marcescens Nuclease having a Chitin Binding Domain tag at its C-terminus and with the poly-histidine tag on the C-terminal side of the CBD-tag.
Dependencies are inferred from the claim text.