Compositions and methods of using a synthetic Dnase I
AssigneeAmbion, Inc.
InventorsGary Latham, Jon Kemppainen
Life of this patent
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Abstract
Compositions and method for making and using a synthetic bovine DNase I are disclosed. More particularly, the sbDNase I of the present invention is a versatile enzyme that cleaves DNA nonspecifically to release 5′-phosphorylated nucleotides. The sbDNase I molecules of the present invention find particular use in a wide range of molecular biology applications, including: degradation of contaminating DNA after RNA isolation; RNA clean-up prior to, or in conjunction with, RT-PCR after in vitro transcription; identification of protein binding sequences on DNA (DNase I footprinting); prevention of clumping when handling cultured cells; tissue dissociation and creation of fragmented DNA for in vitro recombination reactions.
Claims23
An isolated and purified nucleic acid comprising a nucleic acid encoding the protein of SEQ ID NO:
- 2 for a DNase I.
An expression vector comprising a nucleic acid encoding the protein of SEQ ID NO:
- 2 for a DNase I operably linked to a promoter recognized by a host cell transformed with the vector.
An isolated and purified nucleic acid comprising a nucleic acid sequence encoding a protein of at least about a 95% identity with a nucleic acid sequence encoding the protein of SEQ ID NO:
- 2 for a DNase I.
A host cell transformed with an expression vector comprising a nucleic acid encoding an amino acid sequence of SEQ ID NO:
- 2 for a synthetic bovine DNase I.
A process for making a protein with DNase activity comprising the steps of:
- transforming a host cell with an isolated nucleic acid comprising a nucleotide sequence encoding a DNase Protein with at least about an 95% identity with SEQ ID NO:
- 2 for a DNase;
- and
- culturing the host cell under conditions such that the DNase protein is produced by the host cell.
A synthetic bovine DNase I produced by a process comprising:
- culturing a yeast host cell transformed with an expression vector comprising a DNA sequence comprising the nucleotide sequence encoding the synthetic bovine DNase I as shown in FIG. 1 (SEQ ID NO:2),
- expressing the synthetic bovine DNase I in the cultured yeast host cell;
- and
- isolating the synthetic bovine DNase I.
A process for making a synthetic bovine DNase I comprising the steps of:
- transforming a host cell with a nucleic acid molecule that encodes the bovine DNase I comprising an amino acid sequence of SEQ ID NO:
- 2;
- and
- culturing the host cell under conditions in which the bovine DNase I is produced by the host cell.
Dependencies are inferred from the claim text.