摘要
将人红细胞生成素 (h EPO)基因组 DNA(g DNA)与 1.1kb牛 β-乳球蛋白 (BL G)基因的 5′端上游调控序列融合 ,构建了 p CMV- BL G- EPO- b GH poly A(p CBEA)乳腺定位表达载体。该载体与已构建的大鼠乳清酸蛋白 (WAP)基因 5′端上游调控序列和 h EPO融合基因 p WAP- EPO- WAP3′U TR(p WE3′)和 p CMV- WAP- EPO- b GH poly A (p CWEA )经脂质体包裹后 ,通过显微外科方法 ,经乳腺导管注入妊娠中后期家兔乳腺内 ,进行短暂表达。于家兔分娩后 1～ 2 5 d采奶 ,EL ISA检测乳汁中 EPO含量 ,3个载体均获得表达。表达水平从高到低依次为 p CWEA、p CBEA、p WE3′;表达量为 0 .116～ 0 .75 3μg/ L 。
Three mammary-specific expression vectors of pCBEA,pWE3′ and pCWEA were constructed with 1.1 kb bovine BLG gene promoter,0.949 kb WAP gene promoter and 2.4 kb erythropoietin genomic DNA minigene.The difference among the three vectors was that WAP3′ UTR was used as termination signal in pWE3′ and bGH polyA in pCBEA,pCWEA and the CMV enhancer element was cloned into the upstream of BLG,WAP gene promoter in pCBEA,pCWEA.The vectors mixed with liposome were injected into mammary glands of late stage pregnant rabbits through mammary gland teat canal respectively.Milk was collected on 1-25 d from rabbits after parturition.The results of ELISA assays showed that the expression level of pCBEA,pWE3′ and pCWEA was at the range of 0.335-0.680 μg/L,0.051-0.516 μg/L and 0.201-0.753 μg/L in the rabbit milk respectively.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2002年第3期238-240,共3页
Chinese Journal of Veterinary Science
基金
军队医药卫生重点课题 ( 96 Z0 5 0 )
关键词
牛
BLG
hEPO
大鼠
WAP
家兔
乳腺
基因表达
BLG
WAP
human erythropoietin
mammary gland
gene transfer
expression
rabbit
