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.
ABSTRACT
Objective To study the distributions of three single nucleotide polymorphisms (SNPs) in β-arrestin2 (ARRB2) which including rs3786047,rs1045280 and rs2036657 and to elucidate the relationship between these SNPs and response to methadone maintenance treatment (MMT) among heroin-dependent patients of Han ethnicity population in Hunan.Methods Han MMT patients were recruited in four random-chosen MMT clinics from Hunan province.Demographics,history of drug-use and MMT were recorded.ARRB2 SNPs were genotyped to determine the association between SNPs and response to MMT.Results Distributions of the three SNPs were in Hardy-Weinberg equilibrium in both groups (responders vs.non-responders).There was no statistical significance in the distribution frequency of genotype on rs3786047 (x2=0.486 2,P=0.784),rs1045280 (x2=1.591 9,P=0.451) and rs2036657 (x2=1.061 5,P=0.588) in ARRB2 among the responders or the non-responders.Conclusion Associations between the ARRB2 genotypes,rs3786047,rs1045280 and rs2036657,and MMT response in Han MMT patients in Hunan province did not appear.
ABSTRACT
Objective To study the distributions of three single nucleotide polymorphisms (SNPs) in β-arrestin2 (ARRB2) which including rs3786047,rs1045280 and rs2036657 and to elucidate the relationship between these SNPs and response to methadone maintenance treatment (MMT) among heroin-dependent patients of Han ethnicity population in Hunan.Methods Han MMT patients were recruited in four random-chosen MMT clinics from Hunan province.Demographics,history of drug-use and MMT were recorded.ARRB2 SNPs were genotyped to determine the association between SNPs and response to MMT.Results Distributions of the three SNPs were in Hardy-Weinberg equilibrium in both groups (responders vs.non-responders).There was no statistical significance in the distribution frequency of genotype on rs3786047 (x2=0.486 2,P=0.784),rs1045280 (x2=1.591 9,P=0.451) and rs2036657 (x2=1.061 5,P=0.588) in ARRB2 among the responders or the non-responders.Conclusion Associations between the ARRB2 genotypes,rs3786047,rs1045280 and rs2036657,and MMT response in Han MMT patients in Hunan province did not appear.