Literature summary for 2.7.7.48 extracted from

Kumar, D.; Chaudhury, I.; Kar, P.; Das, R.H.
Site-specific cleavage of HCV genomic RNA and its cloned core and NS5B genes by DNAzyme (2009), J. Gastroenterol. Hepatol., 24, 872-878.

Cloned(Commentary)

Cloned (Comment)	Organism
in vitro transcription of NS5B using SP6 RNA polymerase, and recombinant expression of His6-tagged NS5B and of untagged RDRP in Hep-G2 cells. In vitro cleavage of HCV core and NS5B mRNAs by DNAzyme in Hep-G2 cells is assessed by RT-PCR using RNA from cells co-transfected with cloned core or NS5B gene and its respective DNAzyme leading to reduction of NS5B protein activity in NDNAzyme co-transfected cells	Hepacivirus C

Cloned (Comment)

Organism

in vitro transcription of NS5B using SP6 RNA polymerase, and recombinant expression of His6-tagged NS5B and of untagged RDRP in Hep-G2 cells. In vitro cleavage of HCV core and NS5B mRNAs by DNAzyme in Hep-G2 cells is assessed by RT-PCR using RNA from cells co-transfected with cloned core or NS5B gene and its respective DNAzyme leading to reduction of NS5B protein activity in NDNAzyme co-transfected cells

Hepacivirus C

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	reduction of NS5B activity by co-expression of an NS5B mRNA-cleaving DNAzyme in Hep-G2 cells	Hepacivirus C

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	-	Hepacivirus C

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
42000	-	x * 42000, recombinant His6-tagged NS5B, SDS-PAGE, x * 68000, recombinant RDRP, SDS-PAGE	Hepacivirus C
68000	-	x * 42000, recombinant His6-tagged NS5B, SDS-PAGE, x * 68000, recombinant RDRP, SDS-PAGE	Hepacivirus C

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
nucleoside triphosphate + RNAn	Hepacivirus C	-	diphosphate + RNAn+1	-	?

Organism

Organism	UniProt	Comment	Textmining
Hepacivirus C	Q0G9P7	i.e. HCV	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged NS5B from Hep-G2 cells by nickel affinity chromatography	Hepacivirus C

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
nucleoside triphosphate + RNAn	-	Hepacivirus C	diphosphate + RNAn+1	-	?
nucleoside triphosphate + RNAn	nculeotides are GTP, CTP, ATP, and UTP, the RNA templates for the enzyme assay are transcribed from linearized murine inducible nitric oxide synthase, iNOS, clone having 400 nt insert in an in vitro transcription reaction using SP6 RNA polymerase	Hepacivirus C	diphosphate + RNAn+1	-	?

Subunits

Subunits	Comment	Organism
?	x * 42000, recombinant His6-tagged NS5B, SDS-PAGE, x * 68000, recombinant RDRP, SDS-PAGE	Hepacivirus C

Synonyms

Synonyms	Comment	Organism
NS5B	-	Hepacivirus C
RDRP	-	Hepacivirus C
RNA dependent RNA polymerase	-	Hepacivirus C

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Hepacivirus C

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Hepacivirus C