Reverse transcriptase having improved thermostability

一种逆转录酶、热稳定性的技术,应用在转移酶、微生物的测定/检验、酶等方向

Active Publication Date: 2013-10-09
BIONEER
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The present invention is provided to solve the problems of conventional reverse transcriptases as described above

Method used

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  • Reverse transcriptase having improved thermostability
  • Reverse transcriptase having improved thermostability
  • Reverse transcriptase having improved thermostability

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Embodiment Construction

[0023] The present invention is described in detail below.

[0024]The present invention provides a reverse transcriptase having improved thermostability, which is obtained by substituting other amino acid residues for one or more amino acid residues selected from the group consisting of the following amino acid residues: Provided: Glutamine at position 63 (Q63), lysine at position 264 (K264), and position 295 in the amino acid sequence of reverse transcriptase derived from M-MLV (represented by SEQ.ID.NO:1) Lysine (K295), threonine (T306) at position 306, glutamic acid at position 346 (E346), proline at position 408 (P408), histidine at position 438 (H438 ) and asparagine at position 454 (N454).

[0025] Amino acid substitutions herein include one or more substitutions selected from the group consisting of the following substitutions, but are not always limited thereto: substitution of glutamine at position 63 with leucine (Q63L), substitution of glutamine at position 63 wit...

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Abstract

The present invention relates to a reverse transcriptase having improved thermostability, more precisely a mutant reverse transcriptase with improved thermostability by substitution of one or more amino acids selected from the group consisting of the 63rd glutamine (Q63), the 264th lysine (K264), the 295th lysine (K295), the 306th threonine (T306), the 346th glutamic acid (E346), the 408th proline (P408), the 438th histidine (H438), and the 454th asparagin (N454) of the amino acid sequence of M-MLV originated reverse transcriptase represented by SEQ. ID. NO: 1 with other amino acids. The mutant reverse transcriptase of the present invention demonstrates excellent thermostability, compared with the wild type reverse transcriptase. Therefore, it is advantageous to obtain the target cDNA with stable reverse transcription activity even in the presence of RNA that can form the stable secondary structure at a high temperature.

Description

technical field [0001] The present invention relates to reverse transcriptases, and more specifically to reverse transcriptases having improved thermostability obtained by mutating specific amino acid residues that make up conventional reverse transcriptases. Background technique [0002] Much work has been done on the reversal of the origin of RNA tumor viruses, especially Moloney-Murine Leukemia Virus (M-MLV), human HIV, or avian myeloblastosis virus (AMV) Recording enzymes have been studied and their functions and properties have thus been published. Reverse transcriptase has been used in many molecular biology methods such as cDNA library construction, reverse transcription, and polymerase chain reaction (PCR) due to its unique property of facilitating the synthesis of complementary DNA (cDNA) by using RNA as a template. )Wait. [0003] Three prototypes of the reverse transcriptase of retroviruses were mainly studied. Moloney murine leukemia virus-derived retroviruses...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12C12N15/54C12N15/63
CPCC12P19/34C12N9/12C12N9/1276C12N15/52C12Q1/48C12N1/20C12N15/70Y02P20/52
Inventor 朴翰浯梁成俊朱成谟黄炳午
Owner BIONEER
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