Recombinant T4 RNA Ligase 2, truncated K227Q is a truncated form of T4 RNA Ligase 2 recombinantly expressed in Escherichia coli, containing the K227Q mutation.

This enzyme specifically joins the 5'-preadenylated terminus of DNA or RNA to the 3'-OH terminus of RNA. No ATP is consumed during ligation; the reaction is initiated by preadenylated adaptors. The K227Q mutation greatly reduces the enzyme’s inherent weak activity of transferring the adenylyl group from 5'-preadenylated DNA adaptors (AppDNA) to RNA 5'-phosphates to form 5'-phosphorylated DNA adaptors (pDNA), thereby minimizing non-specific RNA ligation such as RNA concatenation or self-circularization.

In addition, this enzyme exclusively uses 5'-preadenylated single-stranded DNA or RNA as 3'-adaptors without ATP consumption, which significantly reduces background in ligation reactions, making it the preferred choice for microRNA cloning and library construction. The enzyme can be inactivated by heat treatment at 65 °C for 20 minutes.

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