Application of gene knockout animal as skin barrier function disorder animal model
A technology of gene knockout and dysfunction, applied in animal husbandry and other fields, can solve the problems of poor uniformity of model preparation standards, many uncontrollable factors, and poor similarity of clinical disease models
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[0029] 1: Construction of transgenic mice
[0030] The inventors utilized the Cre / loxP recombinase system to convert Cdc42 loxp / loxp Transgenic mice were crossed with K5-Cre transgenic mice, and the genotype of the F1 generation mice obtained was Cdc42 loxp / + / -K5-Cre+. Then use Cdc42 loxp / + / -K5-Cre+ mice and Cdc42 loxp / loxp Transgenic mice were crossed, and their offspring obtained 1 / 4 ratio of Cdc42 gene knockout (knockout) mice, referred to as KO mice, and the genotype was Cdc42 loxp / loxp / -K5-Cre+; Obtain 1 / 2 ratio of Cdc42 gene non-knockout (wild type) mice, referred to as WT mice, the genotype is Cdc42 loxp / + / K5-Cre- or Cdc42 loxp / loxp / -K5-Cre-. Build process such as figure 1 shown.
[0031] Gene level and protein level identification were carried out by PCR method and Cdc42 immunohistochemical method.
[0032] Mouse tissue PCR gene identification: K5-Cre+ is a 667bp specific band, Cdc42 loxP / loxP It is a 700bp specific band, WT is a 600bp specific band; K5-...
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