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An artificial expression system and application of insect acetylcholine receptor protein complex

A technology of acetylcholine receptor and expression system, which is applied in the field of artificial expression system of insect acetylcholine receptor protein complex, can solve the problem of not fully reflecting the function of insect acetylcholine receptor and the characteristics of active compounds, and achieve high accuracy and specificity Good sex, good effect of detection and screening platform

Inactive Publication Date: 2018-11-16
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, the so-called "insect acetylcholine receptor" expressed by this method contains mammalian acetylcholine receptor subunits or part of non-insect subunit sequences, and the analysis of active compounds obtained by using this expression system and protein function and The results of the research on the characteristics cannot fully reflect the function of the real insect acetylcholine receptors and the characteristics of the active compounds

Method used

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  • An artificial expression system and application of insect acetylcholine receptor protein complex

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Embodiment 1

[0032] 1. the present embodiment is the source of acetylcholine receptor and TCP1 with East Asian migratory locust, and expresses acetylcholine receptor α1 subunit and β1 subunit and TCP1 with Xenopus laevis oocyte as artificial expression system to illustrate specific implementation, but this Methods are not limited to the materials and expression systems described in this example. At the same time, a control experiment was carried out using the Xenopus laevis oocyte artificial expression system. Control experiment 1 is the currently used insect acetylcholine receptor expression method, specifically expressing only the α1 subunit (Locα1) of the migratory locust acetylcholine receptor and the β2 subunit (rβ2) of the rat; For the α1 subunit and β1 subunit of the locust acetylcholine receptor, 3 control experiments only expressed the TCP1 protein complex (α-θ subunit). The control experiment is not a necessary experiment for this method, and it is here to prove that this method ...

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Abstract

The invention discloses an artificial expressing system for insect acetylcholine receptor protein complexes and an application thereof. As insect acetylcholine receptors and TCP1 are jointly expressed in the artificial system, in-vitro recombination of all functional insect subunit acetylcholine receptors can be achieved; the characteristics of active compounds can be detected and analyzed through the system, and the artificial expressing system and the application are used for detecting and developing pesticide and the like.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to an artificial expression system and application of insect acetylcholine receptor protein complex. Background technique [0002] Nicotinic acetylcholine receptors are important functional proteins located on the cell membrane and belong to the cysteine ​​ring ligand-gated ion channel superfamily. Mediates rapid cholinergic activity in the nervous system and at the junction of nerves and muscles. Acetylcholine receptor is a transmembrane protein complex composed of five subunits distributed on the cell surface, and the five subunits are arranged in a ring to form a central ion channel. Each subunit contains 4 transmembrane segments, namely M1-M4, in which M1, M3 and M4 are β-sheets, M2 is a curved α-helix in the middle, and M2 of the 5 subunits forms the inner lining of the channel. They are like a molecular switch. When combined with an agonist such as acetylcholine, the confo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N15/12G01N33/68
Inventor 刘泽文鲍海波张懿熙
Owner NANJING AGRICULTURAL UNIVERSITY
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