摘要
目的建立表达PiggyBac转座酶转基因小鼠模型,为研究PiggyBac转座子介导基因修饰在小鼠中的应用提供工具。方法利用Cytomegalovirus(CMV)启动子驱动PiggyBac转座酶基因的表达,经显微注射法建立C57BL/6J表达PiggyBac转座酶的转基因小鼠。PCR鉴定转基因小鼠的基因型,RT-PCR检测PiggyBac转座酶在小鼠生殖系睾丸中的表达情况。PiggyBac转座酶转基因小鼠活性的检测,是通过与转座子供体转基因小鼠杂交检测供体位置变化来确定的。结果显微注射产生7只转基因小鼠并能传代,经RT-PCR筛选出一株在睾丸中相对高表达PiggyBac转座酶的转基因小鼠。随后与转座子供体转基因小鼠杂交,子代双阳小鼠与野生型小鼠杂交基因型分离,产生的子代转座子供体单阳性小鼠中具有转座子供体片段的转座反应。结论成功建立了表达Piggy-Bac转座酶转基因小鼠动物模型,该模型为PiggyBac转座子技术在小鼠中的应用提供了有价值的工具动物。
Objective To establish a PiggyBac transposase-expressing transgenic mouse model for study of the genetic modification mediated by transposon in mice.Methods PiggyBac transposase gene was driven by CMV promoter and transgenic mice were created by the microinjection.The gene type of transgenic line was identified by PCR and the gene expression level in testis was determined by RT-PCR.The activity of the PiggyBac transposase was detected with the transposition efficiency by Southern blot in the mice crossed with transposon donor transgenic mice.Results Seven lines of transposase transgenic mice were obtained by microinjection.One mouse line with relatively higher expression level of transposase in the testis was obtained and the transposase induced the transposition of the transposon donor DNA fragment in the mouse genome.Conclusion A PiggyBac transposase-expressing transgenic mouse model is successfully established.This model will greatly contribute to the research of the genetic modification mediated by transposon in mice.
出处
《中国实验动物学报》
CAS
CSCD
2012年第3期60-64,共5页
Acta Laboratorium Animalis Scientia Sinica
基金
协和青年基金
基本科研业务费(DWS201011)
科技重大专项课题"啮齿类研发平台创新药物研究开发技术平台建设"(2011ZX09307-302)
作者简介
马元武（1983-），男，博士生，研究方向：分子遗传学。E—mail：mayuanwu@gmail．com
[通信作者】张连峰。E-mail：Zhanglf@cnilas．org
