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Laminaria gamma-type carbonic anhydrase gene sjγ-ca and its encoded protein and application

A technology of carbonic anhydrase and protein coding, which is applied in the field of genetic engineering to achieve the effect of increasing production

Active Publication Date: 2022-05-24
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Accounting for about 90% of the total inorganic carbon content of seawater, it is the main form of seawater inorganic carbon; while CO 2 The proportion is less than 1% of the total inorganic carbon, but the photosynthetic efficiency of large algae such as kelp is much higher than that of land plants (Gao & McKinley, 1994)

Method used

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  • Laminaria gamma-type carbonic anhydrase gene sjγ-ca and its encoded protein and application
  • Laminaria gamma-type carbonic anhydrase gene sjγ-ca and its encoded protein and application
  • Laminaria gamma-type carbonic anhydrase gene sjγ-ca and its encoded protein and application

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Embodiment Construction

[0030] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0031] 1. Materials

[0032]The kelp gametophyte material is self-made, and the temperature is 17±1℃ and the light intensity is 40μmol photons / (m 2 s) and the light / dark ratio of 16h / 8h. The medium used was PES (Starr & Zeikus, 1993), and the medium was changed every two weeks.

[0033] E. coli cloned strain DH5α, expression strain BL21(DE3) and enhanced HRP-DAB substrate chromogenic kit were purchased from Tiangen Biochemical Technology Co., Ltd.

[0034] The expression vector pET28a was purchased from Dingguo Biological Company.

[0035] Taq DNA polymerase, restriction endonucleases BamHI and NotI, T4 DNA ligase, TA cloning vector pMD19-T were purchased from TaKaRa Company.

[0036] Bio-Scale TM Mini Profinity TM IMAC Cartridges protein affinity chromatography prepacked columns were purchased from Bio-Rad Company.

[0037] TA cloning vector pTO...

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Abstract

The present invention discloses kelp γ-type carbonic anhydrase gene Sjγ-CA and its encoded protein and its application. The cDNA sequence of Sjγ-CA is shown in SEQ ID NO: 1, the DNA sequence is shown in SEQ ID NO: 2, and its encoded protein The amino acid sequence of rSjγ-CA is shown in SEQ ID NO:3, and the amino acid sequence of the recombinant protein rSjγ-CA obtained through the construction of the above-mentioned gene Sjγ-CA and its expression vector is shown in SEQ ID NO:4, and has CO 2 Enzyme activity of hydration reaction; the above-mentioned gene Sjγ-CA of the present invention, its encoded protein and recombinant protein rSjγ-CA can be used to cultivate high-yield transgenic kelp crops.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a kelp γ-type carbonic anhydrase gene Sjγ-CA and its encoded protein and application. Background technique [0002] Kelp (Saccharina japonica) to CO 2 The absorption of water helps to mitigate greenhouse gases and acidification of water bodies. It accounts for about 90% of the total inorganic carbon content of seawater, and is the main form of seawater inorganic carbon; while CO 2 It accounts for less than 1% of the total inorganic carbon, but the photosynthetic efficiency of macroalgae such as kelp is much higher than that of land plants (Gao & McKinley, 1994). This is because most algae have an inorganic carbon concentration mechanism (CCM) that increases CO around ribulose 1,5-diphosphate carboxylase / oxygenase (RuBisCO) 2 concentration, so that the algae can fix more CO 2 to improve its photosynthesis efficiency. In CCM, carbonic anhydrase (CA) is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/60C12N9/88
CPCC12N9/88C12Y402/01001Y02P60/20
Inventor 周志刚许玲毕燕会
Owner SHANGHAI OCEAN UNIV