Construction method and application of dcf1-knocked-out mouse model

A dcf1, mouse model technology, applied in the use of vectors to introduce foreign genetic material, cells modified by introducing foreign genetic material, recombinant DNA technology, etc. Effect

Inactive Publication Date: 2015-11-11
SHANGHAI UNIV
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  • Abstract
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Problems solved by technology

[0004] The existing mouse models of olfactory disorder are basically constructed by artificially destroying the olfactory nerve or killing the olfactory mucosal cells with chemical substanc

Method used

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  • Construction method and application of dcf1-knocked-out mouse model
  • Construction method and application of dcf1-knocked-out mouse model
  • Construction method and application of dcf1-knocked-out mouse model

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Experimental program
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Embodiment 1

[0032] dcf1 knockout mouse construction method, see figure 1 .

[0033] 1. The mouse dcf1 gene is located on chromosome 4, and the mouse dcf1 gene is cloned from the mouse BAC library (ID: RP23-423N6). The construction of the targeting vector pLNT-dcf1 is based on the reconstruction of the pLNT plasmid as the parent plasmid, such as figure 1 As shown, three genes are inserted in the middle, and the DNA fragment of the first exon (Exon1) of the dcf1 gene is inserted between BamHI and BamHI, and two 1oxP sites arranged in the same direction are installed on both sides, and the neo gene is positive Screening marker gene, PGK-TK gene is a negative screening marker gene, also has 1oxP sites on both sides. The length of the upstream homologous recombination arm (F1) is 1.2kb (NotI—SalI), the length of the downstream homologous recombination arm (F3) is 4.8kb (ClaI—KpnI), and the length of the intermediate condition deletion part (F2) is 1.4kb, which is dcf1 Part of the first exon...

Embodiment 2

[0042] Example 2: Detection of olfactory sensitivity of dcf1 knockout mice by food burial bead test

[0043]On the day before the experiment, the mice were forbidden to eat, and drinking water was provided normally. Rat food was not provided and the rats were left in a state of extreme hunger. Each cage was covered with 3 cm thick corncob litter, and a grain of 0.5 g rat food was randomly buried in each cage, with a depth of about 0.5 cm. Experimenter 1 puts the mouse into the test cage and covers the cage cover (to prevent the mouse from climbing out of the cage). Experimenter 2 starts timing at the same time. After the mouse finds the mouse food and digs it out, grabs it with its forelimbs. The experiment is over and stops. Time and record the time. If the mouse is not found within 5 minutes, the experiment will also end. The control group included 10 wild-type mice and 10 dcf1 knockout mice each. Each mouse corresponds to a cage, and it is guaranteed that there will be n...

Embodiment 3

[0046] The olfactory preference assay detects olfactory preference in dcf1 knockout mice

[0047] The experimental mice were individually placed in each cage for 1 hour to acclimate to the environment. Aim the camera at one side of the cage from the side, put a 3cm×3cm filter paper in the cage, and soak the filter paper with the specified solution. The experiment time was 3 minutes, the experiment was ended, the filter paper was taken out, and the mice were put back into the original cage. The control group included 10 wild-type mice and 10 dcf1 knockout mice each. A stopwatch was used to count the total time the mice sniffed the filter paper. If the mouse licked the filter paper with its tongue or grasped the filter paper with its forelimbs, these times could not be counted.

[0048] Mice can distinguish between different odors, which can be roughly divided into three categories: preferred odors, neutral odors (neither like nor dislike) and aversive odors. In the olfactory...

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Abstract

The invention relates to a construction method and an application of a dcf1-knocked-out mouse model and relates to an application of olfaction researching of the animal model. In the invention, by means of food-buried pellet test, olfaction preference test and odor avoiding test, sensitivity, judgment and preference of olfaction of the dcf1-knocked-out mouse is carried out. A result proves that the dcf1-knocked-out mouse is reduced in sensitivity and judgment of olfaction and is changed in preference of olfaction. Compared with that a dysosmia mouse model in the prior art requires manual injury or chemical injury to establish the model, wherein the method wastes time and energy and is liable to cause other injuries of mouse, the dcf1-knocked-out mouse model is free of these steps, is stable in dysosmia character and is a suitable mouse model for researching the olfaction.

Description

technical field [0001] The invention relates to a method for constructing a dcf1 knockout mouse model and its application. Background technique [0002] The sense of smell, like vision and hearing, is one of the primitive sensory functions of the human body. The sense of smell can affect people's emotions through the central nervous system. Patients with olfactory disorders have decreased interest in surrounding things, flat reactions, and even serious mental depression. Therefore, the basic research on the sense of smell has very important practical significance. [0003] Mouse models are one of the most commonly used research tools in biological research, and are widely used in the construction and research of various disease models. There is also a long history of research on olfactory function in mice, which is generally conducted by constructing a mouse model of olfactory impairment. [0004] The existing mouse models of olfactory disorder are basically constructed by...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N5/10A01K67/027
Inventor 文铁桥郭健健冯瑞丽王娇
Owner SHANGHAI UNIV
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