摘要
人凝血ＩＸ因子基因乳腺组织特异性表达载体的构建及其在奶山羊乳腺中的分泌性表达张克忠卢大儒邱信芳（复旦大学遗传学研究所上海２００４３３）黄英黄淑帧（上海市儿童医院医学遗传研究所上海２０００４０）血友病Ｂ（ＨｅｍｏｐｈｉｌｉａＢ）是由于人凝血ＩＸ因子（ｈ...
Two expression vectors comprising of mouse matrix attachment regions(MARs),bovine β casein gene sequence,human factor IX(hFIX) minigene and cDNA,pMCIXm and pMCIX,were constructed for the purpose of mammary gland bioreactor.An secretory expression system of hFIX protein in milk was made by using stearylamine(SA)liposome to transfect plasmid DNA into mammary gland lobule of lactating goat directly.The highest production of hFIX in goat milk was 13 7ng/ml 3 days after transfection,and the hFIX production in the goat mammary gland tansfected with pMCIXm containing hFIX minigene was obviously higher than that of transfected with pMCIX containing hFIX cDNA.Activity immuno analysis and barium citrate absorption method showed that>90% hFIX protein in milk appeared to be γ glycosylated and biological activity.This result confirmed the validity of the constructed vectors for further transgenic study,and this assay could also find its success in the evaluation of a foreign gene expression and secretion in the milk as a rapid detective system by using SA liposome to transfect DNA into goat mammary gland directly.
出处
《生物工程学报》
CAS
CSCD
北大核心
1997年第4期419-422,共4页
Chinese Journal of Biotechnology
基金
国家高技术研究项目
上海市科委科技攻关项目
