CXCL4基因敲除小鼠的饲养、繁殖及基因型鉴定

Breeding,reproduction and genotype identification of CXCL4 knockout mice

目的饲养、繁殖并鉴定CXC趋化因子配体4[chemokine(C-X-C motif)ligand 4,CXCL4]基因敲除小鼠,为深入研究CXCL4的生物学功能奠定基础。方法将引进的杂合子小鼠进行饲养并繁殖,剪取繁殖成功的子代小鼠耳朵,提取其基因组DNA,采用PCR法鉴定小鼠的基因型;比较野生型小鼠和CXCL4敲除的纯合子小鼠的体重变化及结直肠的长度,通过HE染色观察野生型小鼠和CXCL4敲除的纯合子小鼠的结直肠形态结构;采用ELISA法验证CXCL4野生型和纯合子小鼠血清中CXCL4的表达。结果繁殖成功的子代小鼠有3种基因型:野生型、杂合子及纯合子;4、10周龄雌性纯合子小鼠体重明显重于同龄的野生型小鼠(P<0.05),6、10周龄雄性纯合子小鼠体重明显重于同龄的野生型小鼠(P<0.05);6周龄雌性纯合子小鼠结直肠长度明显长于同周龄的野生型小鼠(P<0.05),但结直肠的形态结构无显著改变;纯合子小鼠血清中CXCL4的表达量明显低于WT小鼠(P<0.05)。结论成功获得了CXCL4基因敲除纯合子小鼠,为深入研究CXCL4的生物学功能奠定了基础。

Objective To breed,reproduce and identify chemokine（C-X-C motif）ligand 4（CXCL4）knockout mice and lay a foundation of further study on biological function of CXCL4.Methods The introduced heterozygote mice were bred and reproduced,and the ears of offspring mice were collected,from which genomic DNA was extracted and identified for genotype by PCR.The bodyweights and lengths of colorectal of wild type and CXCL4 knockout homozygote mice were compared,while the morphological structures were observed by HE staining.The expression of CXCL4 in sera of wild type and homozygote mice were determined by ELISA.Results The reproduced offspring mice were of three genotypes,i.e.wild type,homozygote and heterozygote.The bodyweights of female homozygote mice at ages of 4 and 10 weeks and male homozygote mice at ages of 6 and 10 weeks were significantly higher than those of wild type mice at the corresponding ages（each P〈.05）.However,the colorectal of female homozygote mice at age of 6 weeks was significantly longer than that of wild type mice at the same age（P〈.05）,of which the morphological structure showed no significant change.The expression level of CXCL4 in sera of homozygote mice was significantly lower than that of wild type mice（P〈.05）.Conclusion CXCL4 knockout homozygote mice were successfully obtained,which laid a foundation of further study on the biological function of CXCL4.