Selective inactivation of M-MuLV RT RNase H activity by site-directed PEGylation: an improved ability to synthesize long cDNA molecules.
N Biotechnol
; 29(3): 285-92, 2012 Feb 15.
Article
in En
| MEDLINE
| ID: mdl-21807127
Moloney murine leukemia virus reverse transcriptase (M-MuLV RT) is a domain structured enzyme that has the N-terminally located DNA polymerization activity and C-terminally located RNase H activity, which interferes with the efficient synthesis of long cDNA molecules. Here we present the PEGylation as a tool for engineering the M-MuLV RT derivative deficient in RNase H activity. We demonstrate that site-directed chemical modification (SDCM) of the RNase H domain by selectively PEGylating C635, one of the eight cysteine residues present in the reverse transcriptase (RT), specifically inactivated its ribonucleolytic activity. As a consequence, the efficiency of long cDNA molecules synthesis by modified enzyme was greatly increased.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Polyethylene Glycols
/
RNA-Directed DNA Polymerase
/
DNA, Complementary
/
Ribonuclease H
/
Reverse Transcription
/
Moloney murine leukemia virus
Limits:
Animals
Language:
En
Journal:
N Biotechnol
Journal subject:
BIOLOGIA MOLECULAR
/
ENGENHARIA BIOMEDICA
Year:
2012
Document type:
Article
Affiliation country:
Lithuania
Country of publication:
Netherlands