Artificially modified high-activity Mariner-Like transposase

A transposase, transposon technology, applied in the field of molecular biology, can solve the problem of loss of catalytic transposition ability, low activity or inactivity, affecting the application of MLE transposon, etc.

Inactive Publication Date: 2014-02-26
ZHEJIANG FORESTRY UNIVERSITY
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Problems solved by technology

However, the MLE transposase isolated in nature has accumulated more or less mutations due to the "vertical inactivation" effect during the evolution process, and has partially or completely lost the ability to catalyze transposition, becoming a low-activity or inactive transposase. It affects the application of MLE transposons, so it is very important to artificially construct highly active transposases

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  • Artificially modified high-activity Mariner-Like transposase
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  • Artificially modified high-activity Mariner-Like transposase

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Embodiment Construction

[0014] The present invention will be further described below in conjunction with specific implementation steps. It should be understood that these examples are only for illustrating the present invention and are not intended to limit the scope of the present invention. Unless otherwise specified, the experimental methods in the following examples without specific conditions are usually operated according to 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.

[0015] 1. Acquisition of wild-type MLE transposase and non-autonomous transposons without transposase

[0016] Using fresh leaves of Moso bamboo (Phyllostachyspubescens, collected in the Botanical Garden of Zhejiang A&F University, N30°15′14.67″N latitude N30°15′14.67″E119°43′33.47″E) as materials, the Genomic DNA of Moso bamboo was extracted by CTAB method, and designed according to the conserved se...

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Abstract

The invention discloses artificially modified high-activity Mariner-Like transposase. Active transposase cloned from moso bamboo and plant MLE (Mariner-Like Elements) transposase with higher activity obtained after molecular optimization of the active transposase lay a foundation for developing genetic labels by means of the MLE transposase and provide a powerful guarantee for large-scale separation and marking of gene functions in the gene function.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to several highly active Mariner-Like transposases. 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 plant genes. [0003] Mariner-Like Transposons (Mariner-Like Elements, MLE) are an important family of transposons, which were first discovered when studying an unstable mutation in the wh...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/90C12N15/61
CPCC12N9/1241
Inventor 周明兵汤定钦郑丽娜
Owner ZHEJIANG FORESTRY UNIVERSITY
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