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We hypothesized that the thermal stability of Moloney murine leukaemia virus reverse transcriptase (MMLV RT) will increase with increases in its ability to bind with a template–primer (T/P). To test this hypothesis, we introduced positive charges into MMLV RT by site-directed mutagensis at positions that have been implicated in the interaction with T/P. Thirty-six variants were constructed in which...
We have previously indicated that three single mutations (Leu144→Ser, Asp150→Glu, and Ile168→Ala) in the site-directed mutagenesis of thermolysin increase the activity and two single (Ser53→Asp and Leu155→Ala) and one triple (Gly8→Cys/Asn60→Cys/Ser65→Pro) mutations increase the stability. In the present study, aiming to generate highly active and stable thermolysin variants, we combined these mutations...
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