Enhancing thermostability of Moloney murine leukemia virus reverse transcriptase through greedy combination of multiple mutant residues.

ABSTRACT: Reverse transcription is crucial in bioengineering and biomedical fields, particularly for genome sequencing and virus diagnosis. Enhancing the thermostability of reverse transcriptase can significantly improve its efficiency and accuracy by enabling it to function at higher temperatures, thereby reducing RNA secondary structures and minimizing interference from contaminating enzymes, particularly in clinical samples. Here, using a combinatorial strategy, a variant of Moloney Murine Leukemia Virus reverse transcriptase (MMLV RT) with improved activity across a wide temperature range (30-50 °C) was identified and maintained 100% activity after incubation at 50 °C for 10 min. Eleven hot-spot residues were mutated in various combinations, and the mutant proteins were rapidly expressed in a ce

SUBMITTER: Yang Y 

PROVIDER: S-EPMC11842686 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature