Construction and identification of a CDHR2 gene conditional knockout mouse model
10.12025/j.issn.1008-6358.2017.20170245
- VernacularTitle:CDHR2基因条件性敲除小鼠模型的构建及鉴定
- Author:
Zi-Yuan XIA
1
;
Hua-Bo JIANG
;
Yang WANG
;
Mei-Jin HUANG
;
Bin LU
Author Information
1. 第二军医大学药学院生化药学教研室
- Keywords:
CDHR2;
genknockout;
Cre/loxP system
- From:
Chinese Journal of Clinical Medicine
2017;24(2):176-180
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a CDHR2 conditional knockout mouse model, and to provide conditions for the study on biological function of CDHR2 gene.Methods:The conditional CDHR2 targeting vector was constructed and transfected into mouse embryonic stem (ES) cells by electroporation.The positive ES cells screened by G418 and GANC were microinjected into the blastocysts of C57BL/6J mice.The chimeric mice were obtained and then mated with Cre mice to obtain conditional CDHR2 knockout mice.The phenotype was analyzed by PCR and immunohistochemistry, respectively.Results:The conditional CDHR2 targeting vector was successfully constructed, with six positive clones of ES cells being obtained.The positive clones of ES cells were microinjected into blastocysts of C57BL/6J mice with 5 chimeric mice being obtained.The chimeric mice were mated with Flp mice, and 6 positive F1 generation mice without Neo gene were obtained.Finally, such mice were hybridized with Cre mice to obtain intestine-specific CDHR2 knockout mice.Immunohistochemistry assay showed that the CDHR2 gene in intestinal tracts of the positive mice was specifically knocked out.In contrast, the expression of CDHR2 in kidney tissue was not affected.Conclusions:The successful construction of intestine-specific CDHR2 knockout mice has laid the foundation for provides a basis for further functional study on CDHR2 gene.
