Modified RNA ligase for efficient 3′ modification of RNA

摘要

The invention provides a novel truncated mutated T4 RNA ligase 2. In addition, methods are provided for ligating pre-adenlylated donor molecules to the 3′ hydroxyl group of RNA in the absence of ATP using the ligase.

主权项

1. A method for enzymatically ligating a pre-adenylated donor molecule to RNA, the method comprising reacting the pre-adenylated donor molecule and the 3′ hydroxyl group of a 5′ phosphorylated or de-phosphorylated RNA in the absence of adenosine triphosphate and in the presence of an enzyme comprising a truncated T4 RNA ligase 2 (1-249),
wherein said enzyme includes a substitution of a lysine with an arginine at a position corresponding to position 225 as encoded by SEQ ID. NO: 3, and wherein the truncated T4 RNA ligase 2 is capable of ligating the pre-adenylated donor molecule to the 3′ hydroxyl group of the optionally de-phosphorylated RNA in the absence of adenosine triphosphate.