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Artificial nuclease and preparation method thereof

A hydrolase, nucleic acid technology, applied in the field of biochemistry, to achieve the effect of efficient cutting

Inactive Publication Date: 2012-04-25
乔元彪 +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The technical problem to be solved by the present invention is to provide a kind of artificial nuclease and its preparation method according to the restriction relationship between the solubility, molecular structure, size and activity of the existing artificial nuclease

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  • Artificial nuclease and preparation method thereof
  • Artificial nuclease and preparation method thereof
  • Artificial nuclease and preparation method thereof

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Embodiment Construction

[0035] The preparation method of artificial nucleic acid hydrolase comprises the following steps:

[0036] (1) Preparation of artificial nucleic acid hydrolase Ⅰ: Stir glycine (or glycine), 1,10-phenanthroline and copper chloride (or copper hydroxide) in the water phase at a molar ratio of 1:1:1 Mix, react at 55°C for 30 minutes, filter after cooling to room temperature;

[0037] (2) Preparation of artificial nucleic acid hydrolase II: Stir and mix glycylglycine, 1-methylimidazole and newly prepared copper hydroxide (or copper chloride) in the water phase at a molar ratio of 1:1:1, and heat to React at 55°C for 30 minutes, filter after cooling;

[0038] (3) After mixing the filtrate prepared in step (1) and step (2) with N,N-dimethylformamide at a volume ratio of 1:3, the solution appears turbid, and then add water drop by drop Turn into a transparent solution, add 3-5 drops of water again and let it stand for volatilization. After the product of step (1) stands for volatili...

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Abstract

The invention discloses an artificial nuclease and a preparation method thereof, belonging to the technical field of biochemistry and aiming at providing an efficient artificial nuclease and a preparation method thereof according to a restriction relation among the solubility, molecular structure, size and activity of the traditional nuclease. A water soluble functional metal compound is prepared from copper chloride, glycine and pi-electronic groups in a water phase through one-step chemical reaction, is of a plane or tetragonal pyramidal structure, and has strong insertion and hydrolysis capacity on nucleic acid, and stronger sequence selectivity and connection capacity; and by using a mixed solvent volatilizing method, the crystallizing efficiency of products is quickened, and the purity and the yield are improved. Copper at the center of the compound obtained from the invention is respectively of a three-dimensional tetragonal pyramidal structure and a plane quadrilateral structure, thus the artificial nuclease is endowed with tRNA targeting identification and biological connection genes, and has stronger binding constant to the nucleic acid and high hydrolysis rate.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and specifically relates to an artificial nucleolytic enzyme, and also relates to a preparation method of the artificial nucleic acid hydrolase. Background technique [0002] Phosphodiester bonds in nucleic acids are fairly stable under physiological and non-enzymatic conditions. The widely used restriction endonuclease recognition sites are generally a few nucleotides, and they can only break nucleic acids at specific sites, so their use is limited. Chemically synthesized DNA / RNA cleavage compounds are internationally developed hydrolysis tools, which can effectively attack and hydrolyze phosphodiester bonds and generate 5'-, 3'-oligonucleoside phosphates. Artificial nucleases can effectively remove the contamination of pathogenic microorganisms such as bacteria and viruses; in addition, they have very important application value in modern molecular biology such as antisense nucleic acid, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/22C12N15/55
Inventor 乔元彪杨卫民吴玉花
Owner 乔元彪
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