摘要
为研究人 DAF基因在小鼠体内遗传与表达的规律 ,从质粒 p SFFV- DAF分离出一段包含人 DAF基因的 DNA片段 .采用受精卵显微注射法建立转人 DAF基因小鼠 .提取出生小鼠的染色体 DNA,经 Dot- blot与 Southern- blot杂交相结合确定首建转基因小鼠 ,并经Dot- blot杂交研究人 DAF基因在转基因小鼠体内的遗传特征 ,Northern杂交确定其表达情况 .小鼠受精卵经基因导入后 ,共生出 2 4只小鼠 ,其中 4只被确定为首建转基因小鼠 ,整合率为 1 5% .在首建转基因小鼠两两交配生出的 F1代小鼠中分别有 70 %和 75%继续携带人DAF基因 .首建转基因小鼠中有 1只小鼠在 RNA水平表达了人 DAF基因 .可见 ,人 DAF基因整合入小鼠基因组中 ,并能够稳定遗传及表达 .
To study the regulation of integration and expression for hDAF gene in transgenic mice,a DNA fragment contains hDAF gene from pSFFV DAF was isolated and then injected into the fertilized eggs of mice using pronucleus microinjection approach. When newborn mice are 4 weeks old, the genomic DNA was isolated from their tail. Dot blot and Southern blot hybridization were performed to identify the founder transgenic mice. Dot blot hybridization was used to analyse intergration of hDAF gene in the offspring from founder mice, and Northern blot hybridization was used to test expression of hDAF gene in transgenic mice.24 mice were born after microinjection.Four of them were ascertained as transgenic mice for hDAF gene, and the intergration frequency was 17%. There are respectively 70% and 75% of offspring mice from founder mice were integrated with hDAF gene. hDAF gene can be expressed at RNA level in kidney of 1 transgenic mouse. It shows hDAF gene integrated into genomic DNA of mice, and can be heredited and expressed in transgenic mice.
出处
《生命科学研究》
CAS
CSCD
2000年第4期321-327,共7页
Life Science Research
基金
国家自然科学基金项目!( 39570 671 7)
