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3.1.26.13: retroviral ribonuclease H

This is an abbreviated version!
For detailed information about retroviral ribonuclease H, go to the full flat file.

Word Map on EC 3.1.26.13

Reaction

Endohydrolysis of RNA in RNA/DNA hybrids. Three different cleavage modes: 1. sequence-specific internal cleavage of RNA. Human immunodeficiency virus type 1 and Moloney murine leukemia virus enzymes prefer to cleave the RNA strand one nucleotide away from the RNA-DNA junction. 2. RNA 5'-end directed cleavage 13-19 nucleotides from the RNA end. 3. DNA 3'-end directed cleavage 15-20 nucleotides away from the primer terminus. =

Synonyms

HIV reverse transcriptase ribonuclease H, HIV reverse transcriptase-associated RNase H, HIV RNase H, HIV RT RNaseH, HIV RT-associated RNase H, HIV-1 reverse transcriptase, HIV-1 reverse transcriptase ribonuclease H, HIV-1 reverse transcriptase RNase H, HIV-1 RH, HIV-1 ribonuclease H, HIV-1 ribonuclease H domain, HIV-1 RNase H, HIV-1 RT-associated RNase H, human immunodeficiency virus reverse transcriptase, human immunodeficiency virus reverse transcriptase-associated ribonuclease H, human immunodeficiency virus reverse transcriptase-associated RNase H, human immunodeficiency virus RT-associated RNase H, M-MuLV RT RNase H, reverse transcriptase, reverse transcriptase-associated ribonuclease H, reverse transcriptase-associated RNase H, reverse-transcriptase-associated ribonuclease H, ribonuclease H, ribonuclease H domain in HIV-1 reverse transcriptase, RNase H, RNase H activity of HIV-1 reverse transcriptase, RNase H domain of HIV-1 reverse transcriptase, RNase H of HIV-1 subtype C, RNaseH, RNH, RNHHIV, RT RNase H, RT-RNase H

ECTree

     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.26 Endoribonucleases producing 5'-phosphomonoesters
                3.1.26.13 retroviral ribonuclease H

Cloned

Cloned on EC 3.1.26.13 - retroviral ribonuclease H

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
cloning a His-tagged p66 or p51 HIV-1 RT DNA fragment into the pQE-9 vector and expression in Escherichia coli strain BL21(DE3)pLys
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expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli XL1 Blue cells
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expression in Escherichia coli
expression of His-tagged enzyme
expression of His6-tagged p66/p51 HIV-1 reverse transcriptase p51/p66 52A variant C280S, the p66 subunit also contains the mutations K172A and K173A
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expression of RNase H domain from residue Y427 to L560 as fusion protein in Escherichia coli
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expression of RNase H, in an N-terminally His-tagged construct designed to encode the p51 subunit joined by a linker to the thumb (T), connection (C), and RNase H (R) domains of p66, in Escherichia coli strain MIC2067(DE3) lacking endogenous RNase HI and HII as soluble protein. The construct G provides sufficient RNase H activity to complement Escherichia coli growth at 42°C
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expression of the isolated RNase H domain in Escherichia coli
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isolated 125-residue RNase H domain consisiting of residues G436-L560 of polypepitde p6
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recombinant expression of both enzyme subunits, N-terminally His-tagged p66 and p51 subunit mutants C282S of HIV-1, in Escherichia coli strain BL21 Rosetta
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recombinant expression of enzyme mutant C280S
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recombinant expression of His-tagged enzyme in Escherichia coli strain M15
Human immunodeficiency virus type 1 group M subtype B
recombinant expression of mutant constructs
recombinant expression of wild-type and mutant RNase H RNH domains (residues 427-560) in Escherichia coli strain Rosetta 2 (DE3) as N-terminally His6-SUMO-fused proteins
recombinant overexpression of wild-type enzyme and chimeric mutants in Escherichia coli strain MIC2067 (DE3)
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separate expression of subunits in Escherichia coli
the entire RNase H region of reverse transcriptase RT is encoded in codons 431-560, DNA and amino acid sequence determination and analysis, genotyping
Human immunodeficiency virus type 1 subtype C
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transfection of HEK 293 cells
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