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Extraction and detection method of phytase in phytase transgenic corn kernels

An extraction method and a phytase transfer technology, which are applied in the field of bioengineering and can solve problems such as the absence of phytase extraction and detection methods.

Inactive Publication Date: 2011-07-20
JILIN ACAD OF AGRI SCI
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  • Application Information

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Problems solved by technology

However, in the analysis of existing patents and documents, it is found that there are no patents and document reports on the extraction and detection methods of phytase in corn grains with phytase genes

Method used

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  • Extraction and detection method of phytase in phytase transgenic corn kernels
  • Extraction and detection method of phytase in phytase transgenic corn kernels
  • Extraction and detection method of phytase in phytase transgenic corn kernels

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Embodiment Construction

[0018] Below in conjunction with specific embodiment, further illustrate the present invention. This embodiment is only used to illustrate the present invention and is not intended to limit the scope of the present invention. The experimental methods that do not indicate specific conditions in the following examples are usually operated under conventional conditions.

[0019] One experimental material

[0020] The phytase gene-transferred corn and its recipient corn are planted in the National Transgenic Plant Pilot Test and Industrialization Base (Gongzhuling), and the mature corn grains are collected from the phytase gene-transferred corn and its recipient corn, pulverized by a grinder, and placed in a Store in -70°C refrigerator.

[0021] Second reagent

[0022] Phytase standard product (30U / g) was purchased from Sigma Company, and sodium phytate, anhydrous sodium acetate, ammonium molybdate, and ammonium metavanadate were all purchased from BBI Company. Potassium dihyd...

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Abstract

The invention discloses an extraction and detection method of phytase in phytase transgenic corn kernels, which is characterized in that: pH of extract is 5.50, a solid-liquid ratio is 1:4, and extraction time is 15min; and the extract, and 0.1mmol / L substrate application solution react in acetic acid buffer solution with a pH of 5.50 for 30min. The invention has the advantages that: the extraction method of the phytase in the phytase transgenic corn kernels is optimized, optimization experiments are performed on the basis of the standards of 'Feed Phytase Activity Determination-Spectrophotometry', and the best method for the extraction and determination of the phytase in the phytase transgenic corn kernels is determined so as to provide related data for constructing a phytase transgenic corn safety assessment system.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to an extraction and detection method optimization of phytase in transgenic crops, in particular to an extraction and detection method applicable to phytase gene transgenic corn grains. Background technique [0002] Since 1990, when Monsanto and Decca first obtained genetically modified corn, the genetically modified corn industry has achieved great success. In 2009, the global planting area of ​​genetically modified corn was 158 million hectares, accounting for 26% of the global planting area of ​​genetically modified crops. In 2009, the phytase gene-transferred corn obtained the safety certificate for the production and application of agricultural genetically modified organisms officially issued by the Ministry of Agriculture. Phytase (Phytase) belongs to phosphohydrolase, which is a general term for enzymes that hydrolyze phytic acid and its salts in plants into available i...

Claims

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Application Information

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IPC IPC(8): C12N9/16C12Q1/44
Inventor 宋新元于壮刘娜李静张欣芳张明李飞武李葱葱邢珍娟邵改革康岭生董立明夏蔚赵宁
Owner JILIN ACAD OF AGRI SCI
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