目的:建立人CTRP4基因的转基因小鼠,为脂肪细胞因子CTRP4的体内功能研究奠定基础。方法首先构建人CTRP4的转基因小鼠线性化表达载体,再利用显微注射的方法将载体注射入小鼠受精卵,从而构建人CTRP4的首建鼠( Founder )并与野生型小鼠交配繁殖得到F1代阳性小鼠,再通过近亲繁殖与测交的方法,得到CTRP4转基因纯合子小鼠,并通过PCR和western blot 的方法对纯合子小鼠进行鉴定。结果得到人CTRP4转基因小鼠纯合子小鼠两个品系,western blot鉴定该转基因小鼠心脏,肝脏,脑,肾脏等多种组织中均呈现CTRP4高表达。结论成功构建了人CTRP4转基因小鼠纯合子小鼠。
Objective To establish the CTRP4 transgenic mouse model and investigate the function of the novel adipocytokine CTRP4.Methods CTRP4 overexpressing vector in pCAGGS was firstly constructed and then microinjected into zygote to establish the founder transgenic mice .F1 heterozygotes were generated by founder mice mating with wildtype mice, and the CTRP4 transgenic homozygotes were generated by F 1 littermates.The genotype was confirmed by PCR and test cross method .The expression level of CTRP 4 in transgenic mice was detected by western blot .Result The human CTRP4 transgenic homozygote mice line was established , and the expression level of CTRP 4 was confirmed raletively high in detected tissues including heart , liver, brain and kidney . Conclusion The human CTRP4 transgenic mice was successfully established .