Spyda基因过表达转基因小鼠品系的建立  

Identification of transgenic mice with Spyda gene over expression

作　　者：张瑶楠[1] 丁宁[1] 石心泉[1] 贾孟春[1] 刘美玲[1] 

机构地区：[1]国家人口计生委科学技术研究所,国家卫生计生委男性生殖健康重点实验室,北京100081

出　　处：《生殖医学杂志》2014年第10期838-842,共5页Journal of Reproductive Medicine

基　　金：国家自然科学基金资助项目(31201116;30670784)

摘　　要：目的建立Spyda基因过表达转基因小鼠品系用于深入研究Spyda基因的功能。方法利用Gateway技术构建Spyda表达载体,通过DNA显微注射的方式获得Spyda基因过表达的首建鼠。首建鼠与野生型鼠杂交,所得子代中的阳性鼠同窝交配,已传3代以后的阳性小鼠经SYBR Green实时荧光定量PCR方法检测外源基因的起始模板量,通过与杂合子比较来预测小鼠的基因型,再用传统育种方式验证SYBR Green实时荧光定量PCR的结果。结果获得7只首建鼠,通过与野生型鼠杂交获得阳性子代,进行同窝交配,传至第3代,经实时荧光定量PCR方法筛选出4只纯合子的小鼠,经测交验证,其与正常小鼠交配所生的后代均为阳性,证明实时定量PCR的结果是正确的。建立了2个独立的Spyda转基因小鼠纯合子品系。结论建立了稳定遗传的纯合子Spyda基因过表达转基因小鼠品系,可作为工具鼠进一步深入研究Spyda基因的功能。Objective： To establish transgenic mice with Spyda gene over-expression to study the function of Spyda gene. Methods： The mice with Spyda gene over-expression were obtained by microinjecting the Spyda expression vector which was established by Gateway technology. After three generations, the levels of original template detected by real-time fluorescent quantitative PCR were compared between the heterozygous and homozygous mice and the results were validated by using cross test. Results： Seven Spyda gene over-expression mice were obtained, and four homozygous mice were screened by real-time fluorescent quantitative PCR. The results of cross test indicated that the offspring of the homozygous mice mated with normal mice were all positive. These were consistent with the results of real-time fluorescent quantitative PCR. Two independent homozygous transgenic mouse strains had been established. Conclusions： Spyda gene over-expression transgenic mouse strains had been established. The hornozygous transgenic mouse is a tool to study the function of Spyda gene.

关 键 词：Spyda基因 转基因小鼠 实时荧光定量PCR 纯合子 

分 类 号：R7[医药卫生—临床医学]