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Heat-proof alkaline phosphatase (Tthxm-AP) gene and use of encoding protein thereof

An encoding and heat-resistant technology, which is applied in the field of application of the heat-resistant alkaline phosphatase (Tthxm-AP) gene and its encoded protein, can solve problems such as poor thermal stability

Inactive Publication Date: 2008-09-03
THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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Problems solved by technology

[0003] At present, the most commonly used alkaline phosphatases are calf intestinal alkaline phosphatase and Escherichia coli alkaline phosphatase. Although they have the characteristics of high specific activity, their thermal stability is poor, which limits the application of enzymes in some fields.

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  • Heat-proof alkaline phosphatase (Tthxm-AP) gene and use of encoding protein thereof
  • Heat-proof alkaline phosphatase (Tthxm-AP) gene and use of encoding protein thereof
  • Heat-proof alkaline phosphatase (Tthxm-AP) gene and use of encoding protein thereof

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Embodiment Construction

[0026] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. For the experimental methods not indicated in the following examples, the experimental conditions described in the Molecular Cloning Laboratory Handbook (New York: Cold Spring Harbor Laboratory Press, 2001) are usually followed by conventional conditions, or according to the recommendations of the reagent or instrument manufacturer conditions of.

[0027] In order to achieve the above object, the present invention adopts the following technical measures, and its concrete steps are:

[0028] 1. Extraction of Thermus thermophilus XM Genomic DNA

[0029]Use an inoculation loop to pick the Thermus thermophilus XM strains preserved at -80°C, and put them on the LB medium plate (a kind of bacterial culture medium containing 1% tr...

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Abstract

The invention clones a heat resistant alkaline phosphatase gene from a high-temperature bacteria Thermus thermophilus XM genome. Active polypeptide (enzyme) or modified enzyme modified in genetic engineer which is encoded by the gene can be widely used in research field of diagnostics, amynology, biological chemistry and molecular biology. The invention firstly relates to extraction of Thermus thermophilus XM genome DNA, secondly PCR amplification and sequence analysis of heat resistant alkaline phosphatase gene (tap gene), thirdly expression and purification of heat resistant alkaline phosphatase, fourthly measurement to enzymology property of recombination of alkaline phosphatase.

Description

technical field [0001] The invention relates to a nucleotide sequence of an alkaline phosphatase gene from a high-temperature bacterium (Thermus thermophilus XM) and an amino acid sequence encoded by the gene. The present invention also relates to the protein function encoded by the nucleotide sequence and its application. Background technique [0002] Alkaline phosphatase (alkaline phosphatase) (EC 3.1.3.1) is a non-specific phosphomonoesterase that can catalyze the hydrolysis of almost all phosphate monoesters to produce inorganic phosphate and corresponding alcohols, phenols or sugars. It can also catalyze the transfer reaction of the phosphoryl group, the phosphate group is transferred from the phosphate to the phosphate acceptor such as alcohol, phenol or sugar. Alkaline phosphatase plays an extremely important role in the biogeochemical cycle of phosphorus. Alkaline phosphatase exists widely in various organisms, and the presence of alkaline phosphatase has been dete...

Claims

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Application Information

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IPC IPC(8): C12N15/55C12N9/14C12P21/02
Inventor 徐丽美李建波杨丰
Owner THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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