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A mutant of ppmar1 transposase v376a with high catalytic activity and its application

A V376A, high catalytic activity technology, applied in the biological field, can solve the problems of no catalytic activity and low catalytic activity of MLE transposase

Inactive Publication Date: 2020-09-11
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a Ppmar1 transposase V376A mutant with high catalytic activity and its application, which solves the problem that the MLE transposase isolated from nature has low or no catalytic activity.

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  • A mutant of ppmar1 transposase v376a with high catalytic activity and its application
  • A mutant of ppmar1 transposase v376a with high catalytic activity and its application
  • A mutant of ppmar1 transposase v376a with high catalytic activity and its application

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Embodiment Construction

[0013] The present invention will be described in detail below in conjunction with specific embodiments, but it should be understood that the protection scope of the present invention is not limited by specific embodiments. For the test methods that do not indicate specific conditions in the following examples, they are usually operated under conventional conditions, such as the conditions described in the "Molecular Cloning Experiment Guide" edited by Sambrook et al., or according to the steps stated in the kit, because they do not involve the invention. , so the steps are not described in detail.

[0014] When the examples give numerical ranges, it should be understood that, unless otherwise stated in the present invention, the two endpoints of each numerical range and any value between the two endpoints can be selected. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. ...

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Abstract

The invention discloses a high-catalytic-activity Ppmar1 transposase V376A mutant. An amino acid sequence of the Ppmar1 transposase V376A mutant is as shown in SEQ ID NO.1, and a gene coding nucleotide sequence of the Ppmar1 transposase V376A mutant is as shown in SEQ ID NO.2. Valine at the position 376 of wild type Ppmar1 transposase is mutated into alanine. Activity of the Ppmar1 transposase V376A mutant is 2.59 times of that of the wild type transposase in catalysis of transposon transposition, the foundation is laid for application of MLE (mariner-like elements) transposons for development of gene tagging, and a novel implement is provided for large-scale gene separating and marking and gene function research in the post-genome era.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Ppmar1 transposase V376A mutant with high catalytic activity and application thereof. Background technique [0002] A transposon is a DNA sequence that can be transferred from one site to another on the genome. Since the American geneticist McClintock first discovered transposons (Ac / Ds) in maize in the 1940s, scientists have discovered many types of transposons, which are widely found in bacteria, yeast and higher animals and plants. With the continuous deepening of people's understanding of the structure and function of transposons at the molecular level, some transposons have been transformed into gene tags for gene analysis, and have gradually become one of the important means of large-scale isolation of genes. [0003] Mariner-Like Transposons (Mariner-Like Elements, MLE) are an important family of transposons, which were first discovered when studying an unstable...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/12C12N15/54C12N15/81C12N1/19
CPCC12N9/1241C12N15/81
Inventor 周明兵汤定钦
Owner ZHEJIANG FORESTRY UNIVERSITY