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精子特异性Sleeping Beauty转座酶转基因小鼠模型的建立

Establishment of sperm specific Sleeping Beauty transposase-expressing transgenic mouse
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摘要 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。 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.
作者 路迎冬 张旭 马婧 张连峰 马元武
机构地区 中国医学科学院
出处 《中国比较医学杂志》 CAS 2014年第3期34-38,I0006,共6页 Chinese Journal of Comparative Medicine
基金 协和青年基金(2012J25) 国家重大科技项目(2012BA139B02)
关键词 DNA转座子 SleepingBeauty 转座酶 Sleeping Beauty DNA transposon Sleeping Beauty Transposase
分类号 R332 [医药卫生—人体生理学]
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中国比较医学杂志
2014年 第3期

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