摘要
将氯霉素乙酰转移酶 ( CAT)基因与鸡卵清蛋白基因上游调控序列融合 ,构建了 p CAT-3-OVP(鸡卵清蛋白基因 5′调控序列 )表达载体。将构建的载体与脂质体混合后制备转染液 ,转染鸡原代输卵管上皮细胞和鸡成纤维细胞进行瞬时表达。结果表明 ,在鸡原代输卵管上皮细胞 ,转染后 4 8h表达量最高 ,为 0 .67μg/L;而在鸡成纤维细胞 ,不论有无类固醇激素存在 ,均不表达。结果提示 ,在输卵管细胞中存在细胞特异性转录因子。
The 1.1 kb region of the cloned fragment,consisting of the flanking regulatory regions was combined with the chloramphenicol acetyltransferase reporter gene(CAT gene) and an expression vector,pCAT 3 OVP was successfully constructed.pCAT 3 OVP containing this fusion gene was introduced using lipofectin method into cultured primary chicken oviduct cells and fibroblasts cells of chicken for transient expression of CAT,in which the ovalbumine gene was induced by four kinds of steroid hormones.It was demonstrated that in the primary cell cultures of chicken oviduct,the CAT gene was not expressed in 18 hours before induction with insulin,estradiol and corticosterone.Expression of the gene started 12 hours after induction with these hormones and reached a peak(0.67 μg/L) after 48 hours.In the chicken fibroblasts cell cultures,however,the CAT gene was not expressed both in the presence or absence of steroid hormones.It is suggested that there are some of cell specific transcription factors in the primary cultured chicken oviduct cells.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2001年第1期21-24,共4页
Chinese Journal of Veterinary Science
基金
国家"8 6 3"基金资助项目 !(Z2 1-0 4-0 3)
