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Mutant CPD photo-repair enzyme and 6-4 photo-repair enzyme double-plasmid co-expression bacterial strain and application thereof

A photorepair enzyme and co-expression technology, applied in the field of biology, can solve problems such as hindering DNA replication and transcription

Active Publication Date: 2019-02-22
THE FIRST INST OF OCEANOGRAPHY SOA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If these two pyrimidine dimers are not repaired, DNA replication and transcription are hindered

Method used

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  • Mutant CPD photo-repair enzyme and 6-4 photo-repair enzyme double-plasmid co-expression bacterial strain and application thereof
  • Mutant CPD photo-repair enzyme and 6-4 photo-repair enzyme double-plasmid co-expression bacterial strain and application thereof
  • Mutant CPD photo-repair enzyme and 6-4 photo-repair enzyme double-plasmid co-expression bacterial strain and application thereof

Examples

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Embodiment 1

[0034] The genes of CPD photorepair enzyme and 6-4 photorepair enzyme in this example come from Antarctic ice algae (Chlamydomonassp. ICE-L). The invention provides CPD photorepair enzyme (GenBank accession number: KC344405) and 6-4 photorepair enzyme (GenBank accession number: MF784511) and their expression vectors are BL21(DE3) / pET-28a(Kan+) / CPD photorepair enzyme respectively Repair enzyme and BL21(DE3) / pET-21a(Amp+) / 6-4 photorepair enzyme.

[0035] 1. Acquisition of cDNA sequences of CPD photorepair enzyme and 6-4 photorepair enzyme of Antarctic ice algae

[0036] 1.1. Cultivation of Antarctic ice algae

[0037] The Antarctic ice algae used in this example was separated and purified from seawater and sea ice samples collected near Zhongshan Station (69 ° S, 77 ° E) during China's eighteenth Antarctic scientific expedition in November 2001. It is preserved in the Antarctic ice algae species bank of the Key Laboratory of Marine Bioactive Substances of the State Oceanic Adm...

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Abstract

The invention provides a mutant CPD photo-repair enzyme, a 6-4 photo-repair enzyme double-plasmid co-expression bacterial strain and application thereof. The co-expression strain is obtained by transforming plasmids of the CPD photo-repair enzyme into the 6-4 photo-repair enzyme expression strain and then mutating a glutamic acid on the site 323 of the CPD photo-repair enzyme into alanine. The mutant CPD photo-repair enzyme and the 6-4 photo-repair enzyme double-plasmid co-expression strain are constructed; and the bacterial strain has the ability of the CPD photo-repair enzyme and a 6-4 photo-repair enzyme to effectively repair the DNA damage caused by ultraviolet radiation. The bacterial strain obtained by the site-directed mutagenesis of the CPD photo-repair enzyme significantly improves the ability of resistance to ultraviolet radiation, and is of great significance for studying the effect of UV-B on living organisms and the life mechanism of the living organisms to adapt to UV-B enhancement.

Description

technical field [0001] The invention belongs to the technical field of biology, and in particular relates to a mutant CPD photorepair enzyme and 6-4 photorepair enzyme double-plasmid co-expression strain and application thereof. Background technique [0002] The ozone layer can absorb more than 99% of ultraviolet rays, so that humans and other organisms on the earth will not be damaged by strong solar ultraviolet radiation, so the ozone layer is also called the "protective umbrella" of the earth. However, this "umbrella" has been severely damaged, and extreme weather may also lead to more severe ozone decline in the polar regions. The main effect of the destruction of the ozone layer is the enhancement of UV-B reaching the surface, and UV-B will have adverse effects on animals and plants, and it will be fatal to single-celled organisms such as microalgae. Studies have found that if the ozone in the stratosphere is reduced by 10%, the global incidence of cataracts will incre...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12N15/60C12R1/19
CPCC12N9/88C12N15/70C12Y401/99003Y02A50/30
Inventor 缪锦来曲长凤安美玲郑洲
Owner THE FIRST INST OF OCEANOGRAPHY SOA
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