Literature summary for 2.7.7.49 extracted from

Vichier-Guerre, S.; Ferris, S.; Auberger, N.; Mahiddine, K.; Jestin, J.
A population of thermostable reverse transcriptases evolved from Thermus aquaticus DNA polymerase I by phage display (2006), Angew. Chem., 45, 6133-6137.

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Cloned(Commentary)

Cloned (Comment)	Organism
expression of A608T/E520G/W827R, M747K/E742K, and M761T/D547G/I584V in Escherichia coli	Thermus aquaticus

Protein Variants

Protein Variants	Comment	Organism
A608T/E520G/W827R	substitution rates per base for RNA-dependent DNA is similar or lower than that of avian myeloblastosis virus reverse transcriptase. Rate constants kcat of about two orders of magnitude larger than those of the Stoffel fragment	Thermus aquaticus
M747K/E742K	substitution rates per base for RNA-dependent DNA is similar to that of avian myeloblastosis virus reverse transcriptase	Thermus aquaticus
M761T/D547G/I584V	fidelity which is about 10-times higher than that of avian myeloblastosis virus reverse transcriptase. Almost tenfold improved catalytic efficiency as measured by the kcat/Km ratio when compared with the Stoffel fragment	Thermus aquaticus
additional information	directed polymerase-evolution experiment yields variants endowed with RNA-dependent DNA polymerization. To further estimate the fidelity during reverse transcription, the variants A608T/E520G/W827R, M747K/E742K, and M761T/D547G/I584V are used to copy a part of messenger RNA(mRNA) into complementary DNA (cDNA) prior to subcloning for sequencing of reverse transcription products. The sequences showsd that selected variants can polymerize more than 300 nucleotides and are not limited to the addition of single nucleotides at the 3 end of DNA primers. The substitution rates per base for RNA-dependent DNA polymerization of the most active variants, A608T/E520G/W827R and M747K/E742K are similar or lower than that of avian myeloblastosis virus reverse transcriptase, which is used as a standard. The most abundant variant and M761T/D547G/I584V has a fidelity which is about 10-times higher than that of avian myeloblastosis virus reverse transcriptase	Thermus aquaticus

Organism

Organism	UniProt	Comment	Textmining
Thermus aquaticus	-	-	-

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Release 2023.1 (January 2023)