摘要
以小鼠MAR（matrixattachmentregion）元件，牛β－酪蛋白（β－casein）基因5＇端2．0kb调控顺序，人凝血Ⅸ因子小基因（hFIXminigene）构建乳腺组织特异性表达载体，表达载体和质粒pSV－neo共转染中国仓鼠卵巢细胞（CHO），兔皮肤纤维细胞（RSF）和人胚肾上皮细胞（293细胞）．发现hFIX基因在CHO细胞中获得低水平表达，最高表达量为7．2ng／ml．在皮肤成纤维细胞中的表达量低于2ng／ml．在293细胞中没有表达．表达载体用stearylamine（SA）脂质体包埋后尾静脉注射哺乳期小鼠，hFIX蛋白在小鼠乳汁中表达水平高达87．34ng／ml．
Mammary gland specific expression vector comprising of mouse matrix attachment regions (MARs),a 2. 0 kb bovine β-casein gene sequence, human clotting factor Ⅸ (hFIX)minigene was constructed. Chinese hamster ovary cells (CHO),rabbit skin fibroblast cells (RSF) and human embryo kidney epithelium cells (293 cells) were respectively cotransfected by the expression vector and plasmid pSV-neo. The expression of hFIX gene maintained a low level in CHO cells and the highest expression level in CHO cell supermatant was 7. 2 ng/ml, the expression level of hFIX gene in RSF cell supernat was lower than 2 ng/ml, and the hFIX gene was not able to express in 293 cells. The expression vector packaged by stearylamine (SA) liposome was transferred into lactating mice via tail vein, and the highest expression level of hFIX gene in mouse milk was 87. 34 ng/ml.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
1998年第4期527-531,共5页
Journal of Fudan University：Natural Science
基金
国家科委八六三高科技项目
上海市科委科技攻关项目
关键词
人凝血Ⅸ因子
乳腺组织
基因表达
载体
β-casein gene
human clotting factor Ⅸ
mammary gland expression
