扬州大学学报（农业与生命科学版）2011，Vol.32Issue(3)：6-10,5.
人乳铁蛋白乳腺特异性表达载体的构建及其功能的验证
Construction of mammary gland specific vector for expression of human lactoferrin
摘要
Abstract
To screen transgenic donor cells for nuclear transfer, the cassette of LoxP-cre-GFP-Neo was ligated to 3' end SV40 polyA sequence of pbCNCS/LF36 thus producing Paplm. Fertilized mouse eggs (ICR) were microinjected with the expression cassette of pbCNCS/LF36 and transferred into pseudo-pregnant females. The pregnancy rate of pseu-dopregnancy mices were 28. 5% and, 32 and 43 founder mice were born respectively. 4 (3-♀ , 1 ♂ ) transgenic mice were positive checked by PCR and the integration rate were 12. 5%. F1 mice from male were not transgenic. The expression level of Hlf were 3. 8,2. 8,0. 9 mg·Ml-1 in the GO mice and 4. 2, 3. 9,3. 6 mg · Ml-1 in the milk of 3 F1 mice of 4 number GO mice checked by ELISA. Fetal fibroblast cells were transfected with mammary gland specifical expression vector of Paplm and then screened with G418. The single colony was selected and expanded. Genomic DNA were ana-lysed by PCR. 8 lines of fetal fibroblast cells expressed GFP and these transgenic cells would be used to produce Hlf transgenic goat. These results indicated that milk promoter combined with CMV could direct high expression of Hlf Cdna specifically to the mammary gland in the transgenic mice and the dual selectable marker genes of Paplm could be used to produce transngenic donor cells for nuclear transfer.关键词
人乳铁蛋白/双标记基因/双启动子/小鼠/胎儿成纤维细胞/荧光蛋白Key words
human lactoferrin/ dual selectable marker genes/ dual promo tors/ mice/ fetal fibroblast cell/ GFP分类
生物科学引用本文复制引用
袁玉国,安礼友,于宝利,杨廷佳,成勇..人乳铁蛋白乳腺特异性表达载体的构建及其功能的验证[J].扬州大学学报（农业与生命科学版）,2011,32(3):6-10,5.基金项目
国家转基因生物新品种培育重大专项(2009ZX08008-009B) （2009ZX08008-009B）
江苏省科技支撑计划项目(BE2009328) （BE2009328）
国家自然科学基金资助项目(31101871) （31101871）
