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Experiment method for establishing an animal model for studying oral white sponge nevus of the Chinese

An experimental method and animal model technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, cells modified by introducing foreign genetic material, etc., can solve the problems of unclear molecular mechanism and no effective treatment methods, etc. Low cost, short technical cycle and strong specificity

Inactive Publication Date: 2011-06-22
张剑明
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Problems solved by technology

In 2008, through research, we found that the causative gene mutation of Chinese WSN disease occurred on the Keratin 4 gene, the mutation site is 1829G→A, and the protein E520K, thus revealing the pathogenicity of Chinese people's disease for the first time in the world Gene mutation site, but so far, the molecular mechanism of this disease is still unclear, and there is no effective treatment

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  • Experiment method for establishing an animal model for studying oral white sponge nevus of the Chinese
  • Experiment method for establishing an animal model for studying oral white sponge nevus of the Chinese

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Embodiment Construction

[0032] like figure 1 Shown, the implementation steps of the present invention are as follows:

[0033] Firstly, the DNA of WSN patients was extracted by salting out method. Design two pairs of primers to carry out polymerase chain reaction (PCR) on the target gene on the K4 gene in DNA, sequence the amplified K4 gene fragment, and discover the gene mutation site of Chinese WSN patients.

[0034] Then use the point mutation kit of STRATAGENE company to create a mutation G to A at position 1829 of the KRT4 gene of plasmid pCMV6-XL4-KRT4, so that the amino acid is mutated from E to K. PCR and sequencing prove that the mutation is successful. A linker for ligation was constructed and then recombined using restriction endonucleases to generate a recombined KRT4 gene. The KRT4 gene was inserted into the expression vector pcDNA3.1(+ / -) to complete the construction of the K4 transgenic model recombinant plasmid. The recombinant plasmid was transfected into LA795 murine lung cancer ...

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Abstract

An experiment method for establishing an animal model for studying oral white sponge nevus of the Chinese comprises the following steps of: detecting the mutant gene locus of Chinese WSN patients; establishing K4 mutant gene recombination plasmid; transferring the recombinant plasmid; and screening and identifying K4 transgenic animal model. In the invention, research is carried out aiming at WSN mutant gene locus discovered in Chinese families, and thus the method of the invention possesses certain pertinence and uniqueness. The technology provides short periods, low cost, strong singularity, can basically exhibit symptoms of WSN, and is helpful for experiment research on pathogenesis of WSN disease.

Description

Technical field: [0001] The invention relates to an experimental technique applied to the study of human oral mucosal disease-white spongy spot, and transfers the human oral disease to an animal model so as to provide effective experimental animals for the research and treatment of the disease mechanism. Background technique: [0002] Oral white sponge nevus (hereinafter referred to as WSN), also known as white wrinkle disease, is an autosomal dominant genetic disease caused by keratin gene mutation, characterized by irregular penetrance, and the individual incidence rate is related to There are many family series, and the point mutation of proteins K13 and K4 expressed in acanthine cells or the insertion of exogenous bases lead to the occurrence of the disease. In the current research on the mutation gene of white cavernous stains, a total of 9 mutation sites have been found in the world (2 of which were discovered by us). By comparing other mutation sites, we found that th...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/85C12N5/10A01K67/027
Inventor 张剑明
Owner 张剑明
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