Propagation of prdm1 gene knockout mouse and its genotype identification.
- Author:
Xiao-Yun LU
1
;
Chong CHEN
;
Xiu-Ying PAN
;
Ling-Yu ZENG
;
Zhen-Yu LI
;
Xu-Guang SONG
;
Kai-Lin XU
Author Information
1. Affiliated Hospital of Xuzhou Medical College, Jiangsu Province, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Genotype;
Mice;
Mice, Inbred C57BL;
genetics;
Mice, Knockout;
genetics;
Reproduction;
Transcription Factors;
genetics
- From:
Journal of Experimental Hematology
2012;20(4):985-988
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to propagate and identify the prdm1 gene-knockout mice, so as to lay the foundation for studying Blimp-1 protein. Two kinds of transgenic homozygous mice with B6.prdm1(flox/flox) and B6.Lck-Cre were feed and propagated; after successful propagating, the first passage mice were obtained; after the first passage mice were copulated once again, the genotypes were obtained as follows: B6. prdm1(wild/wild). Lck-Cre, B6. prdm1(wild/wild), B6.prdm1(flox/flox). Lck-Cre, B6.prdm1(flox/wild). Lck-Cre, B6.prdm1(flox/flox), B6. prdm1(flox/wild). The genomic DNA of second passage mice was extracted, the Cre and loxp gene fragments were amplified by PCR, then the size of Cre and loxp genomic DNA were detected by agarose gel electrophoresis. The mice with B6.prdm1(flow/flox). Lek-Cre were used as conditionally prdm1-knockout mice, B6.prdm1(flox/wild). Lck-Cre mice, B6.prdm1(flox/flox) and B6 mice were used as controls. The spleen T lymphocytes and B lymphocytes were sorted by using magnetic beads, the blimp-1 target protein was identified by Western blot. The results showed that the two transgenic homozygous mice had the ability to reproduce, and the separation ratio of second passage mice generated from propagation of their offspring cach other meet Mendelian laws, and the prdm1 gene-knockout mice also could successfully obtained. It is concluded that the application of Cre-loxp system may successfully obtain plentiful prdm1 gene-knockout mice.
