摘要
构建了含有ＭＣＫ增强子，β－ａｃｔｉｎ启动子，及ｈＦＩＸ内含子１的３个载体Ｃ１ＮａＭＢＡＩＸ， Ｇ１ＮａＭＢＡｉＩＸ， Ｇ１ＮａＰＡＩＸｉ’ＢＡＭ．转入 ＰＡ３１７和成肌细胞 Ｃ２Ｃ１２细胞后测定ｈＦＩＸ的表达，发现反向构建的Ｇ１ＮａＰＡＩＸｉ’ＢＡＭ表达最高且最稳定，而正向构建的Ｇ１ＮａＭＢＡｉＩＸ的内含子常被剪切，在Ｃ２Ｃ１２细胞中表达不高．进一步将转化细胞肌肉注射Ｃ３Ｈ小鼠，发现载体 Ｇ１ ＮａＰＡＩＸｉ’ ＢＡＭ在小鼠血浆中表达最高达１８０ｎｇ／ｍｌ血浆，持续２０ｄ．
This paper reported the construction of three related retroviral vectors, C1NaMBAIX, G1NaMBAiIX and G1NaPAIXi'BAM, which had MCK enhancers and betaactin promoters in common, differed in their nitron components. The vectors were transferred into PA317 and C2C12 cells and the expression of hFIX was tested. The reverse constructed vector G1NaPAIXi'BAM showed the highest expression and the nitron stably existed, while nitron of direct constructed vector G1NaMBAiIX was spliced in C2C12 cells. Further study in vivo by injection of trans feeted C2C12 cells into C3H mice skeletal muscle revealed that the vector G1NaPAIXi'BAM had the highest hFIX expression (180ng /ml plasma) and lasted for 20 days.
出处
《复旦学报（自然科学版）》
CAS
CSCD
北大核心
1998年第2期117-122,共6页
Journal of Fudan University：Natural Science
基金
国家八六三高科技项目
国家自然科学基金
