Protein composite crystal culture box for in-situ X-ray diffraction

Abstract

The invention discloses a protein composite crystal culture box for in-situ X-ray diffraction and aims to solve the technical problem of possible crystal damages caused by taking out protein crystals from a crystallization chamber to implement X-ray diffraction in the technical field of biology so as to provide effective means for research of protein molecular structures and functions and further intelligent drug development. The composite crystal culture box comprises a plurality of composite crystallization chambers and / or vapor-phase crystallization chambers in array and a magnetic material, the two types of the crystallization chambers are internally provided with porous materials for adsorbing and fixing precipitation liquid or equilibrium liquid, thin gel layers for adsorbing and fixing protein fluid and protein crystals and retaining plates for isolating the protein fluid from the precipitation liquid or equilibrium liquid, the retaining plates are provided with notches or gaps to enable direct contact of the protein fluid with the precipitation liquid or contact by virtue of a vapor phase, and after contact, supersaturation of the protein fluid is realized, and the protein crystals are separated out. In use of the protein composite crystal culture box, taking out the crystals is avoided, and the crystal culture box can be directly and laterally erected on an X-ray diffraction instrument.

Description

technical field [0001] The invention discloses a composite crystal growth box for directly placing protein crystals grown in a solution together with a container in X-rays for in-situ diffraction, and belongs to the field of biotechnology. Background technique [0002] Life science is the science of the 21st century. The life science revolution triggered by the Human Genome Project reached its climax in the structural genome or proteome period in the post-genome era, because genes are only the blueprint of life, and proteins are the realization of life. Proteins are not only the "bricks" that construct living organisms, but also the "molecular machines" that perform life activities. [0003] The function of a protein molecular machine depends on its molecular structure. Protein molecules are very complex, usually composed of tens of thousands of carbon, hydrogen, oxygen, nitrogen, sulfur and other atoms. The molecular structure of protein can be divided into I-level struct...

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Problems solved by technology

However, there is no special high-throughput crystallization plate that can be placed sideways on the market at present; the invention patents "an in-situ diffraction device and diffraction method for protein crystals" (publication number CN108593689A) and "near physiological state biological Diffraction method for macromolecular crystals" (publication number CN108732193A) describes the in-situ diffraction crystallization box invented, but the technical scheme of sticking polymer film on both sides of the double-sided adhesive tape cannot fix the crystal when the box is used on its side, and the double-sided adhesive tape There are physical and chemical compatibility problems for many protein crystallization solutions. In addition, the technical plan does not give a layout plan for the protein solution and its equilibrium solution / precipitation solution, that is, it does not specify which crystallization method to use, such as vapor phase diffusion method, liquid- Liquid diffusion method or liquid mixing method, etc.

When the ordinary crystallization plate is placed sideways, the protein crystals will move, the equilibrium liquid and the protein liquid will move, and the X-rays will change direction after being diffracted by the crystal and be blocked by the side wall of the crystallization chamber. Introduced misinformation leading to incorrect molecular structures of proteins resolved

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