High-temp. resistant mismatch repair enzyme gene, polypeptide coded thereby and preparing method thereof

A mismatch repair, high temperature technology, used in the field of mutation or genetic engineering

Inactive Publication Date: 2003-05-28
HUADA GENE RES & DEV CENT HANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In bacteria and yeast, Muts and Mutl form a repair initiation complex that recognizes mismatched DNA, but this has not been found in humans

Method used

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  • High-temp. resistant mismatch repair enzyme gene, polypeptide coded thereby and preparing method thereof
  • High-temp. resistant mismatch repair enzyme gene, polypeptide coded thereby and preparing method thereof
  • High-temp. resistant mismatch repair enzyme gene, polypeptide coded thereby and preparing method thereof

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Embodiment Construction

[0031] First, the present invention provides an isolated polynucleotide molecule encoding a polypeptide with DNA mismatch repair enzyme activity, the nucleotide molecule is obtained by sequencing and analyzing the whole genome of Tengchong thermophilic anaerobic bacteria, and has The nucleotide sequence of SEQ.ID NO.1, which encodes a polypeptide with a reading frame of 869 amino acids, has a predicted molecular weight of 99056 Daltons.

[0032] The present invention also relates to a recombinant vector comprising the isolated nucleotide molecule of the present invention, and a host cell comprising the recombinant vector. Meanwhile, the present invention includes a method for constructing the recombinant vector and host cell, and a method for producing DNA mismatch repair enzymes by recombinant engineering technology.

[0033] The present invention further provides an isolated DNA mismatch repair enzyme or polypeptide, which is characterized by having the amino acid sequence o...

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Abstract

A process for coding the separated DNA with activity and function equivalent variant and preparing the polypeptide with high-temp. resistant DNA mismatch repair enzyme activity and its function equivalent variant from said DNA by recombination is disclosed. The high-temp. resistant DNA mismatch repair enzyme gene is cloned and separated, which can be used to prepare its transgenic microbe, animal, or plant and recover the enzyme coded by said gene. The amino acid sequence of said polypeptide and the processes for preparing, separating and purifying said polypeptide are also disclosed.

Description

technical field [0001] The present invention relates to mutation or genetic engineering, in particular to a high temperature resistant DNA mismatch repair enzyme gene and a polypeptide encoded by it and a preparation method. Background technique [0002] The MutS gene encodes MutS (DNA mismatch repair enzyme), which is a key component of the DNA mismatch repair system (MMR). The enzyme and other proteins (mutH, mutL or uvrD / mutU) form a mismatch repair complex, which uses the energy of ATP hydrolysis to repair the mismatched base pairs of DNA double-strands and loops formed due to deletions and insertions. The stability of the species' genomic DNA reduces the occurrence of various mutations. [0003] In recent years, the research on human cancer and MMR (mismatch repair system) deficiency has turned to the study of eukaryotic MMR, and the key component of which is the MutS protein. Extensive phylogenetic analysis (RASA, or relative apparent synapomorphy analysis) using the...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C12N15/63C12N15/64
Inventor 汪建李蔚纪振动王鹏王俊田微
Owner HUADA GENE RES & DEV CENT HANGZHOU
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