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Kelp alpha type carbonic anhydrase gene Sjalpha-CA2 and encoding protein and application thereof

A-CA2, carbonic anhydrase technology, applied in the field of genetic engineering to achieve the effect of increasing yield

Active Publication Date: 2019-07-30
SHANGHAI OCEAN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

HCO 3 - It is the main form of inorganic carbon in seawater, accounting for about 90% of the total inorganic carbon in seawater, while CO 2 The proportion is less than 1% of the total inorganic carbon, but the photosynthetic efficiency of large seaweeds such as kelp is much higher than that of land plants (Gao & McKinley, 1994)

Method used

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  • Kelp alpha type carbonic anhydrase gene Sjalpha-CA2 and encoding protein and application thereof
  • Kelp alpha type carbonic anhydrase gene Sjalpha-CA2 and encoding protein and application thereof
  • Kelp alpha type carbonic anhydrase gene Sjalpha-CA2 and encoding protein and application thereof

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Embodiment Construction

[0027] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0028] 1. Materials

[0029] Laminaria gametophyte materials are self-made, at a temperature of 17±1°C and a light intensity of 40 μmol photons / (m 2 s) and light / dark ratio of 16h / 8h, the medium used was PES (Starr & Zeikus, 1993), and the medium was changed every two weeks.

[0030]Escherichia coli clone strain DH5α, expression strain BL21(DE3) and enhanced HRP-DAB substrate chromogenic kit were purchased from Tiangen Biochemical Technology Co., Ltd.

[0031] The expression vector pET32a was purchased from Dingguo Biotechnology Company.

[0032] Taq DNA polymerase, restriction endonucleases BamHI and XhoI, T4 DNA ligase, and TA cloning vector pMD19-T were purchased from TaKaRa Company. Bio-Scale TM Mini Profinity TM IMAC Cartridges protein affinity chromatography purification prepacked column was purchased from Bio-Rad.

[0033] Plasmid extract...

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Abstract

The invention discloses a kelp alpha type carbonic anhydrase gene Sjalpha-CA2 and an encoding protein and application thereof. The cDNA sequence of the gene is shown in SEQ ID NO:1, the DNA sequence of the gene is shown in SEQ ID NO:2, the amino acid sequence of the encoding protein of the gene Sjalpha-CA2 is shown in SEQ ID NO:3, by constructing an expression vector of the gene Sjalpha-CA2, the amino acid sequence of the recombinant protein rSjalpha-CA2 is shown in SEQ ID NO:4, and the recombinant protein rSjalpha-CA2 has the enzyme activity of a CO2 hydration reaction and the esterase activity for esterifying nitrophenyl acetate into p-nitrophenol; the gene Sjalpha-CA2, the encoding gene thereof and the recombinant protein rSjalpha-CA2 can be used for high-yield transgenic kelp cultivation.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to kelp α-type carbonic anhydrase gene Sjα-CA2 and its encoded protein and application. Background technique [0002] Kelp (Saccharina japonica) on CO 2 The absorption of water helps to alleviate greenhouse gas and water acidification. HCO 3 - It is the main form of inorganic carbon in seawater, accounting for about 90% of the total inorganic carbon in seawater, while CO 2 The proportion is less than 1% of the total inorganic carbon, but the photosynthetic efficiency of large seaweeds such as kelp is much higher than that of land plants (Gao & McKinley, 1994). This is because most seaweeds have an inorganic carbon concentrating mechanism (CCM) that increases CO around ribulose 1,5-bisphosphate carboxylase / oxygenase (RuBisCO) 2 concentration, so that algae can fix more CO 2 to increase its photosynthetic efficiency. Carbonic anhydrase (CA) is an indispe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/60C12N9/88C12N15/70C12N15/82
CPCC12N9/88C12N15/70C12N15/8261Y02A40/80
Inventor 周志刚王震乔亚明毕燕会
Owner SHANGHAI OCEAN UNIV
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