CAJ | 학술논문

目的：建立精子特异性表达Sleeping Beauty （ SB）转座酶转基因小鼠模型，为研究SB转座子在小鼠中的应用提供工具。方法克隆精子特异性启动子用以驱动SB转座酶基因的表达，建立精子特异性表达SB转座酶的载体，利用显微注射方法建立以C57BL/6J为背景的精子特异性表达SB转座酶的转基因小鼠。 PCR鉴定首建鼠的基因型，western blot（WB）和免疫组织化学（IHC）检测SB转座酶基因在小鼠生殖腺睾丸中的表达情况，筛选睾丸中高表达SB转座酶的转基因小鼠。结果显微注射方式获得了5只首建小鼠，其中3只能稳定传代，利用WB和IHC成功的筛选出一株在精子中高表达SB转座酶的转基因小鼠。结论成功建立了精子特异性高表达SB转座酶转基因小鼠模型，为将SB转座子作为一种基因工程工具应用于小鼠基因修饰模型的建立提供非常重要的工具资源。
목적：건립정자특이성표체Sleeping Beauty （ SB）전좌매전기인소서모형，위연구SB전좌자재소서중적응용제공공구。방법극륭정자특이성계동자용이구동SB전좌매기인적표체，건립정자특이성표체SB전좌매적재체，이용현미주사방법건립이C57BL/6J위배경적정자특이성표체SB전좌매적전기인소서。 PCR감정수건서적기인형，western blot（WB）화면역조직화학（IHC）검측SB전좌매기인재소서생식선고환중적표체정황，사선고환중고표체SB전좌매적전기인소서。결과현미주사방식획득료5지수건소서，기중3지능은정전대，이용WB화IHC성공적사선출일주재정자중고표체SB전좌매적전기인소서。결론성공건립료정자특이성고표체SB전좌매전기인소서모형，위장SB전좌자작위일충기인공정공구응용우소서기인수식모형적건립제공비상중요적공구자원。
Objective To establish the sperm specific Sleeping Beauty ( SB ) transposase-expression transgenic mouse for the study of the genetic modification mediated by transposon system in mouse .Methods Prm1 promoter was cloned from mouse genomic DNA to drive the expression of SB transposase .The transgenic mice were generated by microinjection .The gene type of transgenic line was identified by PCR .The expressing level in testis was determined by western blot and immunohistochemistry (IHC) staining.Results Five lines of transposase transgenic mice were obtained by microinjection and three can be germline .One mouse line with higher expression level of transposase in the testis was obtained.Conclusion One transgenic mouse model with Sleeping Beauty transposase - expression was successfully established .This model will greatly contribute to the research of genetic modification mediated by transposon in mouse.