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The attachment of a DNA‐binding Sso7d‐like protein improves processivity and resistance to inhibitors of M‐MuLV reverse transcriptase
Author(s) -
Oscorbin Igor P.,
Wong Pei Fong,
Boyarskikh Ulyana A.,
Khrapov Evgeny A.,
Filipenko Maksim L.
Publication year - 2020
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13934
Subject(s) - processivity , reverse transcriptase , dna ligase , dna , dna polymerase , microbiology and biotechnology , chemistry , biology , virology , biochemistry , rna , gene
Reverse transcriptases (RTs) are a standard tool in both fundamental studies and diagnostics. RTs should possess elevated temperature optimum, high thermal stability, processivity and tolerance to contaminants. Here, we constructed a set of chimeric RTs, based on the combination of the Moloney murine leukaemia virus (M‐MuLV) RT and either of two DNA‐binding domains: the DNA‐binding domain of the DNA ligase from Pyrococcus abyssi or the DNA‐binding Sto7d protein from Sulfolobus tokodaii . The processivity and efficiency of cDNA synthesis of the chimeric RT with Sto7d at the C‐end are increased several fold. The attachment of Sto7d enhances the tolerance of M‐MuLV RT to the most common amplification inhibitors: NaCl, urea, guanidinium chloride, formamide, components of human whole blood and human blood plasma. Thus, fusing M‐MuLV RT with an additional domain results in more robust and efficient RTs.