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Goldfish Tgf2 transposase and preparation and preservation methods thereof

A preservation method and technology of transposase are applied in the field of preparation of goldfish Tgf2 transposase, and can solve problems such as unfavorable fermentation, easy contamination of miscellaneous bacteria and the like

Inactive Publication Date: 2018-12-07
SHANGHAI OCEAN UNIV
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  • Abstract
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Problems solved by technology

Strains are susceptible to miscellaneous bacteria in the early growth stage, which is not conducive to mass fermentation

Method used

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  • Goldfish Tgf2 transposase and preparation and preservation methods thereof
  • Goldfish Tgf2 transposase and preparation and preservation methods thereof
  • Goldfish Tgf2 transposase and preparation and preservation methods thereof

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Embodiment Construction

[0060] In order to better explain the technical solutions of the present invention, embodiments of the present invention are described in detail below with reference to the accompanying drawings. The following examples are used to further illustrate the present invention, but should not be construed as fixing or limiting the present invention. Unless otherwise specified, the technical features used in the embodiments can be replaced with other technical features known in the art that have equivalent or similar functions or effects without departing from the inventive concept.

[0061] 1 Materials and methods

[0062] 1.1 Materials

[0063] Materials: Plasmid pET28a(+), Escherichia coli DH5α, BL21(DE3) were all preserved by Shanghai Ocean University. Isopropyl-β-D-thiogalactoside (IPTG), kanamycin sulfate, glycerol, sucrose, mannitol and other reagents were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0064] from Tgf2 (GeneBank accession number: HM146132, 4720bp) (a...

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Abstract

The invention discloses a preparation method of a goldfish Tgf2 transposase. According to preference and degeneracy of escherichia coli for codons, 83 rare codons in the goldfish Tgf2 transposase aresynonymously replaced for recombinant expression and purification. The obtain codon-optimized goldfish Tgf2 transposase becomes mild in fermentation conditions and is also improved in enzyme activity.The production of soluble active goldfish Tgf2 transposase protein provides a selective tool for transgenic fish.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a preparation method and a preservation method of goldfish Tgf2 transposase with mild expression conditions and high activity. Background technique [0002] Transposons are a class of DNA sequences that can move in the genome, and are called "jumping factors". The application of transposons plays an important role in fish genetics and breeding, transgenics, and genetic modification of biological genomes. effect. At present, a large number of studies on DNA transposons have been carried out at home and abroad. The results show that the transposase encoded by transposons with autonomous transposition activity can not only improve the transposition efficiency but also promote the transgene and promoter of cultured fish. Capture research. Tol2 and Tgf2 are known as "Pisces" in DNA transposons, of which Tol2 is the first transposon with autonomous transposition activity disco...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/70C12N9/96C12N9/12
CPCC12N9/1241C12N9/96C12N15/70C12N2800/22
Inventor 邹曙明赵茜郑国栋徐海丽郑寿长
Owner SHANGHAI OCEAN UNIV
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