Breeding and genotype identification of Arrb2 gene knockout mice
10.3969/j.issn.1001-1978.2018.06.027
- VernacularTitle:Arrb2基因敲除小鼠的繁育及基因型鉴定
- Author:
Wu-Yi SUN
1
;
Jia-Chang SUN
;
Xin-Ran LI
;
Wen-Ting PENG
;
Wei WEI
Author Information
1. 安徽医科大学临床药理研究所
- Keywords:
Arrb2;
knockout mice;
genotype identification;
PCR;
breeding;
protein expression
- From:
Chinese Pharmacological Bulletin
2018;34(6):878-881
- CountryChina
- Language:Chinese
-
Abstract:
Aim To explore the optimal way of breeding and genotype identification of Arrb2 knockout mice, and to find a simple and quick polymerase chain reaction ( PCR) method for the genotyping of Arrb2 knockout mice. Methods Breeding homozygote genotype of Arrb2 gene knockout mice were copula-ted with wild-type C57BL/6J mice, and then the heterozygous mating were used for mating. The growth and development of off-spring were observed. The genomic DNA was extracted from the tail of two-week-old mice. PCR was employed to amplify the Arrb2 gene fragment, and electrophoresis was used to present the gene type. Results The breeding and reproducing were successful and three genotype offspring, including wild-type,heterozygous and homozygous knockout mice were obtained. Agarose gel electrophoresis results showed the size of PCR prod-ucts was about 186 bp and 224 bp, which was consistent with the expected target gene fragment, and identified Arrb2 gene knockout mice of different genotypes successfully. Western blot analysis demonstrated the lack ofβ-arrestin2 protein in the major organs from Arrb2 -/ - mice compared with Arrb2 +/ + and Arrb2 +/ - mice. Conclusions It is feasible to obtain the homo-zygous Arrb2 knockout mice by inbreeding heterozygotes. It is simple, rapid and reliable to identify mouse genetype by PCR.
