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Construction method for transgene mouse model of overexpressing Rps23r1 gene

A rps23r1, transgenic mouse technology, applied in the field of transgenic mouse model construction, can solve the problem that the driving ability of exogenous gene expression is not too strong

Inactive Publication Date: 2011-01-12
XIAMEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because the human Thy-1 gene promoter is not too strong in driving the expression of exogenous genes, the overexpressed RPS23R1 protein level in these mice is only equivalent to 1 times the endogenous RPS23R1 expression level

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  • Construction method for transgene mouse model of overexpressing Rps23r1 gene
  • Construction method for transgene mouse model of overexpressing Rps23r1 gene
  • Construction method for transgene mouse model of overexpressing Rps23r1 gene

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Experimental program
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Embodiment Construction

[0024] The construction method of the Rps23r1 gene transgenic mouse model mainly includes:

[0025] (1) Cloning of the Rps23r1 gene fragment:

[0026] Design the 5' end primer XhoI-Myc-5 (5'-gcg ctcgag cccacc atggcatcaatgcagaagctg-3') for the Myc marker with the XhoI restriction endonuclease site, and the XhoI restriction endonuclease enzyme Cutting site, primer XhoI-Rps23r1-3 (5'-gcg ctcgag ttaggggcccgtggcgccggca-3') for the 3' end of the Rps23r1 gene end. Using the genomic DNA of the Rps23r1 transgenic mouse driven by the human Thy-1 gene promoter constructed by the applicant as a template, PCR amplification was performed. The obtained PCR product was verified by purification and sequencing to have the DNA sequence shown in the sequence table, that is, the Rps23r1 gene fragment.

[0027] (2) Cloning the Rps23r1 gene fragment into the MoPrP plasmid to construct the MoPrP.Rps23r1 plasmid vector carrying the Rps23r1 gene:

[0028] The PCR fragment of the obtained Rps23r1 gen...

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Abstract

The invention discloses a construction method for a transgene mouse model of overexpressing an Rps23r1 gene, belonging to transgene models, in particular relating to the construction method for the transgene mouse model of specifically over-expressing the Rps23r1 gene in a brain. A transgene mouse of over-expressing the Rps23r1 gene is obtained through cloning the Rps23r1 gene, constructing a recombinant plasmid and injecting into a germ cell of the mouse. A genome of the transgene mouse of over-expressing the Rps23r1 gene stably integrates the Rps23r1 gene driven by a mouse Prion gene promoter and can be stably inherited to offspring and express an exogenous RPS23R1 protein with an Myc mark in the brain. The Rps23r1 transgene mouse is found not to have abnormal changes in the long-term observation and provides effective means for researching and estimating Rps23r1 physiological and pathologic functions in vivo.

Description

technical field [0001] The invention relates to a transgenic model, in particular to a method for constructing a transgenic mouse model that specifically overexpresses the Rps23r1 gene in the brain. Background technique [0002] Alzheimer's disease (AD) is the most common neurodegenerative disease, which seriously endangers human health. In the brain, extracellular amyloid plaques formed by the accumulation of β-amyloid (Aβ) small molecular polypeptides and neurofibrillary tangles formed by the hyperphosphorylation of microtubule-binding protein tau in nerve cells are two important symptoms of AD. pathological features. A large number of experimental evidences show that the massive generation and aggregation of β-amyloid protein (Aβ) is the key cause of Alzheimer's dementia, and the hyperphosphorylation of tau protein is also involved in the occurrence of some neurodegenerative diseases. Therefore, identifying new gene proteins that can affect the production of β-amyloid (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/63A01K67/027
Inventor 张云武许华曦孙良武黄秀梅陈耀民
Owner XIAMEN UNIV
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