Modified T7 RNA polymerase

JP7878619B1Active Publication Date: 2026-06-23TOSOH CORP
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Patent Information

Authority / Receiving Office
JP · JP
Patent Type
Patents
Current Assignee / Owner
TOSOH CORP
Filing Date
2025-09-24
Publication Date
2026-06-23

AI Technical Summary

Benefits of technology

【0009】 本発明者らは、T7RNAポリメラーゼの熱安定性、夾雑物による反応阻害に対する耐性、生産性、および/または比活性(RNA生産性)を向上させるアミノ酸変異を見出した。すなわち、本開示は以下の態様を提供する。

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Abstract

The object of this disclosure is to provide a modified T7RNA polymerase. The above problem is solved by providing a T7RNA polymerase selected from any of (i) to (iii) below: (i) A T7 RNA polymerase having the amino acid sequence described in Sequence ID No. 1, which includes at least the amino acid substitutions shown in (1) below; (1) The amino acid residue corresponding to the 114th valine residue of Sequence ID No. 1 is replaced with an isoleucine residue, threonine residue, alanine residue, cysteine ​​residue, glutamic acid residue, lysine residue, leucine residue, methionine residue, asparagine residue, glutamine residue, arginine residue, serine residue, or tryptophan residue; (ii) A T7 RNA polymerase having an amino acid sequence described in Sequence ID No. 1, which includes at least the amino acid substitutions shown in (1) above, and further includes one or more substitutions, deletions, insertions, and additions of one or more amino acid residues at one or more positions in addition to the amino acid substitutions shown in (1) above, and which has enzymatic activity; (iii) An amino acid sequence described in Sequence ID No. 1, which has 70% or more identity with the entire amino acid sequence including at least the amino acid substitutions shown in (1) above, provided that the amino acid substitutions shown in (1) above are maintained, and is a T7 RNA polymerase that has enzymatic activity.
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Claims

1. A T7 RNA polymerase having the amino acid sequence described in SEQ ID NOs: 6, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, or 82.

2. A polynucleotide encoding the T7 RNA polymerase according to claim 1.

3. An expression vector comprising the polynucleotide described in claim 2.

4. A transformant obtained by transforming a host with the expression vector described in claim 3.

5. The transformant according to claim 4, wherein the host is Escherichia coli.

6. A method for producing T7 RNA polymerase, comprising the steps of culturing the transformant described in claim 4 to express T7 RNA polymerase, and recovering the polymerase expressed from the obtained culture.

7. A target nucleic acid amplification reagent comprising the T7 RNA polymerase described in claim 1.