The attachment of a DNA-binding Sso7d-like protein improves processivity and resistance to inhibitors of M-MuLV reverse transcriptase.
FEBS Lett
; 594(24): 4338-4356, 2020 12.
Article
em En
| MEDLINE
| ID: mdl-32970841
ABSTRACT
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.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes de Fusão
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DNA Polimerase Dirigida por RNA
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Sulfolobus
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Inibidores da Transcriptase Reversa
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Farmacorresistência Viral
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Proteínas de Ligação a DNA
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Vírus da Leucemia Murina de Moloney
Idioma:
En
Revista:
FEBS Lett
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
Federação Russa