人DAF基因在转基因小鼠中的遗传与表达

为研究人ＤＡＦ基因在小鼠体内遗传与表达的规律，从质粒ｐＳＦＦＶ－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２４只小鼠，其中４只被确定为首建转基因小鼠，整合率为１５％，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。

Intergration and Expression of Human Decay Accelerating Factor Gene in Transgenic Mice

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.